Question 1

Early gene synthesis strategies — pBR322 fusion expression context  In historic recombinant DNA experiments with chemically synthesized gene segments for the two DNA strands, the sequences were inserted into plasmid pBR322 adjacent to which reporter…

Accepted Answer

Introduction / Context: Early cloning and gene synthesis often relied on fusing synthetic coding sequences to strong, well-characterized bacterial promoters and reporter genes to ensure expression and straightforward detection. The lacZ (β-galactosidase) system offered robust expression and simple screening via colorimetric substrates (e.g., X-gal), making it a popular choice alongside plasmids like pBR322 or closely related derivatives. Given Data / Assumptions: Chemically synthesized double-stranded DNA was inserted into a plasmid backbone. Fusion to a reporter facilitated expression and detection. Lac-based systems are historically ubiquitous for such work. Concept / Approach: By placing synthetic sequences adjacent to or in-frame with lacZ control elements, researchers could produce detectable fusion proteins or peptide tags, enabling rapid screening. While pUC vectors later became iconic for blue–white screening, early strategies with pBR322 and lac-derived elements also leveraged β-galactosidase. Among the listed enzymes, β-galactosidase is the canonical reporter used for such fusions in E. coli. Step-by-Step Solution: Recognize the historical use of lacZ as a reporter in E. coli.Associate early plasmid backbones (pBR322 and derivatives) with lac-based expression.Infer that fusion near β-galactosidase enabled convenient detection.Select β-galactosidase from the choices. Verification / Alternative check: Classic cloning literature shows lac promoters and β-galactosidase reporter fusions widely used for expression and screening, justifying this selection. Why Other Options Are Wrong: Galactokinase, glucokinase, and acid phosphatase are not the archetypal E. coli reporters for blue–white screening or fusion detection in these early strategies. Common Pitfalls: Confusing pBR322 with later pUC series; while vectors differ, the lacZ fusion concept remains central. Final Answer: β-galactosidase

Question 2

Promoter choice for expressing HBV surface antigen (HBsAg)  In recombinant vaccine development, which promoter has been widely used for cloning and high-level expression of the hepatitis B virus (HBV) surface antigen in yeast-based systems?

Accepted Answer

Introduction / Context: Recombinant hepatitis B vaccines are produced by expressing HBsAg in host cells to form immunogenic virus-like particles. Yeast (Saccharomyces cerevisiae) emerged as a safe and scalable production system. The choice of promoter in the expression vector determines transcription strength and, consequently, antigen yield for vaccine manufacture. Given Data / Assumptions: HBsAg is expressed heterologously to assemble into particles resembling viral envelopes. Yeast promoters such as ADH1 and GAL1 are common for strong expression. Baculovirus polyhedrin promoter is powerful in insect cells, not yeast. Concept / Approach: Among the listed promoters, the alcohol dehydrogenase 1 (ADH1) promoter is a strong, constitutive yeast promoter historically used to drive HBsAg expression for licensed recombinant vaccines. While other systems (e.g., baculovirus with polyhedrin promoter) can express HBsAg in insect cells, the question focuses on cloning for vaccine production widely associated with yeast, making ADH1 the most appropriate answer here. Step-by-Step Solution: Identify production host: Saccharomyces cerevisiae for licensed recombinant HBsAg.Recall common strong yeast promoters: ADH1 is constitutive and robust.Match promoter to host and product history.Select ADH1 as the correct promoter. Verification / Alternative check: Documentation from recombinant vaccine development describes HBsAg expression under yeast promoters (including ADH1), yielding particulate antigen used in commercial vaccines, corroborating promoter choice. Why Other Options Are Wrong: Lac promoter is bacterial; polyhedrin is for insect cells; T7 promoter requires T7 polymerase (bacterial systems); galactokinase promoter is not the standard for high-level HBsAg vaccine production. Common Pitfalls: Assuming the strongest promoter overall (polyhedrin) is correct without considering host compatibility; promoter choice must match the expression system. Final Answer: Alcohol dehydrogenase 1 (ADH1) promoter

Question 3

Vaccinology — In the licensed recombinant vaccine used against Hepatitis B virus (HBV), the immunogen is produced against which specific HBV antigen to generate protective antibodies?

Accepted Answer

Introduction / Context: The modern Hepatitis B vaccine is a landmark of recombinant DNA technology. Understanding which viral antigen is used as the immunogen clarifies how neutralizing immunity is generated and why the vaccine prevents infection and chronic carriage. Given Data / Assumptions: The Hepatitis B virus (HBV) has several antigens: HBsAg (surface), HBcAg (core), and HBeAg (secreted e antigen). Neutralizing antibodies that block viral entry target the surface antigens exposed on virions. Recombinant technology allows safe production of a single viral protein without whole virus. Concept / Approach: Protection against HBV correlates with anti-HBs antibodies, which neutralize virus by preventing attachment and entry into hepatocytes. Recombinant vaccines express HBsAg (typically in yeast), which self-assembles into non-infectious virus-like particles that strongly stimulate protective antibody responses without exposure to infectious virus. Step-by-Step Solution: Identify the protective target: neutralization requires anti-HBs antibodies.Recall vaccine design: recombinant HBsAg is expressed, purified, and formulated with adjuvant.Link immunity to outcome: anti-HBs ≥ protective threshold indicates immunity.Therefore, the vaccine is synthesized against HBsAg. Verification / Alternative check: Serologic testing after vaccination measures anti-HBs titers; individuals with adequate anti-HBs are protected even if anti-HBc is negative, confirming that surface antigen is the key immunogen. Why Other Options Are Wrong: HBcAg: induces cell-mediated responses but is not the principal neutralizing target for licensed vaccines. HBeAg: a secreted protein; antibodies to HBeAg do not confer sterilizing immunity. “All of the above” is incorrect because current recombinant vaccines focus on HBsAg. HBV polymerase is an internal enzyme and not used as the vaccine antigen. Common Pitfalls: Confusing diagnostic markers (HBcAb, HBeAg) with vaccine antigens. Only anti-HBs reliably indicates vaccine-induced protection. Final Answer: Viral surface antigen (HBsAg).

Question 4

Veterinary virology — Foot-and-mouth disease (FMD) in cloven-hoofed animals is caused by which type of pathogen?

Accepted Answer

Introduction / Context: Foot-and-mouth disease (FMD) is a highly contagious disease affecting cattle, pigs, sheep, goats, and wild ruminants. Identifying the pathogen type is foundational for understanding diagnostics, vaccine strategies, and biosecurity measures. Given Data / Assumptions: FMD outbreaks spread rapidly between susceptible livestock. The etiologic agent is small, non-enveloped, and acid-labile. Control relies on vaccination, movement restrictions, and surveillance. Concept / Approach: FMD is caused by Foot-and-mouth disease virus (FMDV), a positive-sense single-stranded RNA virus (genus Aphthovirus, family Picornaviridae). Its RNA genome encodes structural proteins (VP1–VP4) and non-structural proteins; error-prone replication leads to high diversity, complicating control. Step-by-Step Solution: Determine pathogen family: Picornaviridae members are RNA viruses.Recall FMDV properties: small icosahedral capsid, no envelope, ssRNA(+).Link to epidemiology: rapid spread and antigenic variation align with RNA virus behavior.Hence, the correct category is RNA virus. Verification / Alternative check: Laboratory RT-PCR is the frontline molecular diagnostic, directly indicating an RNA genome; electron microscopy and antigen ELISAs further confirm identity. Why Other Options Are Wrong: DNA virus: inconsistent with picornaviral classification. Bacteria/protozoa/fungi: do not match clinical course, structure, or molecular tests for FMD. Common Pitfalls: Confusing FMD with vesicular diseases of other etiologies (e.g., vesicular stomatitis, a rhabdovirus), which are also RNA viruses but distinct pathogens. Final Answer: RNA virus.

Question 5

Biopharma in India — The first indigenously developed genetically engineered (recombinant) Hepatitis B vaccine in India was produced by which company?

Accepted Answer

Introduction / Context: India’s biotechnology sector made global headlines by developing an affordable recombinant Hepatitis B vaccine. Recognizing the pioneering company helps students connect scientific innovation with public health impact. Given Data / Assumptions: Recombinant HBV vaccines express HBsAg in heterologous hosts. Affordable vaccines improved coverage in national immunization programs. The question asks about the first Indian recombinant HBV vaccine developer. Concept / Approach: Shantha Biotechnics Pvt. Ltd. (later part of Sanofi) launched an indigenous recombinant Hepatitis B vaccine (often known as Shanvac-B). It used recombinant HBsAg expression and helped drive down costs, improving access across India and other low- and middle-income countries. Step-by-Step Solution: Identify product: recombinant HBV vaccine expressing HBsAg.Recall the Indian innovator: Shantha Biotechnics Pvt. Ltd.Link to impact: enhanced affordability and wider immunization.Select Shantha Biotechnics as the correct answer. Verification / Alternative check: Press releases, WHO prequalification records, and Indian biotech histories cite Shantha’s early HBV vaccine as a milestone in domestic biopharma innovation. Why Other Options Are Wrong: Ranbaxy/Dabur/Glaxo Pvt. Ltd.: not credited with the first Indian recombinant HBV vaccine. Biocon: a major biotech company, but not the first to launch an Indian HBV recombinant vaccine. Common Pitfalls: Confusing multinational brand availability with indigenous development and manufacturing firsts. Final Answer: Shantha Biotechnics Pvt. Ltd..

Question 6

Biotechnology milestones — What is widely recognized as the first therapeutic product manufactured using recombinant DNA technology for human medicine?

Accepted Answer

Introduction / Context: The earliest success of recombinant DNA technology in therapeutics transformed diabetes care. Knowing which product came first helps anchor the timeline of biopharmaceutical development and regulatory milestones. Given Data / Assumptions: Recombinant therapeutics require cloning of human genes into microbial hosts. Purity and consistency are superior to animal-sourced products. Regulatory approvals mark historic “firsts.” Concept / Approach: Recombinant human insulin (often called Humulin) was the first FDA-approved therapeutic produced by recombinant DNA technology in the early 1980s. Produced in Escherichia coli (and later in yeast), it replaced animal-derived insulin, reducing immunogenicity and improving supply reliability. Step-by-Step Solution: Identify earliest rDNA therapeutic: recombinant human insulin.Note benefits: human sequence, scalable fermentation, high purity.Contrast with other products: hGH and tPA were approved later; vaccines are prophylactic rather than classic therapeutic drugs.Therefore, insulin is the first rDNA therapeutic. Verification / Alternative check: Biotech histories and regulatory archives consistently cite insulin as the first marketed recombinant therapeutic, setting standards for bioprocessing and quality control. Why Other Options Are Wrong: hGH/tPA: important rDNA products but approved after insulin. HBV vaccine: a recombinant vaccine, not the first recombinant therapeutic drug. FMD vaccine: veterinary and developed later, not a human therapeutic first. Common Pitfalls: Conflating “first recombinant vaccine” with “first recombinant therapeutic drug.” These categories are distinct in biopharma history. Final Answer: Insulin.

Question 7

Recombinant insulin processing — In early E. coli expression strategies, the A and B chains of insulin were expressed as fusion proteins with β-galactosidase. Which reagent was classically used to cleave the fusion at methionine residues to release …

Accepted Answer

Introduction / Context: Early recombinant insulin production in bacteria relied on expressing the A and B chains as part of large, stable fusion proteins (often with β-galactosidase) to enhance expression and protect peptides from degradation. A specific chemical cleavage step was then used to liberate the insulin chains cleanly for refolding and disulfide pairing. Given Data / Assumptions: CNBr cleaves specifically at the carboxyl side of methionine residues in strong acid. Insulin A and B chains were engineered with methionine junctions to β-gal fusions. After cleavage, chains are purified and correctly reoxidized to form native insulin. Concept / Approach: Cyanogen bromide is a classical reagent for site-specific chemical cleavage at methionine. By designing the fusion protein with a methionine between β-galactosidase and the insulin peptide, CNBr treatment releases the desired chain without proteolysis-induced heterogeneity associated with less specific enzymes. Step-by-Step Solution: Engineer fusion with a Met at the junction.Treat inclusion bodies with CNBr in formic acid to cleave at Met.Purify liberated A and B chains.Refold and form the correct disulfide bonds to yield active insulin. Verification / Alternative check: Classical process descriptions for early recombinant insulin consistently mention CNBr cleavage of β-gal fusions, followed by oxidative folding steps to assemble biologically active insulin. Why Other Options Are Wrong: Hydrogen chloride/hydroxylamine: not the standard site-specific approach for Met cleavage. General peptidases/proteases: lack the desired specificity and can over-digest the product. Common Pitfalls: Assuming proteases were preferred; chemical specificity at methionine made CNBr the historical choice in these constructs. Final Answer: Cyanogen bromide (CNBr).

Question 8

Human physiology — Human growth hormone (hGH, somatotropin) is secreted by which endocrine source in the body?

Accepted Answer

Introduction / Context: Human growth hormone (hGH) is central to linear growth, metabolism, and tissue repair. Knowing its source is essential for interpreting endocrine feedback loops and disorders such as gigantism, acromegaly, and growth hormone deficiency. Given Data / Assumptions: Endocrine regulation involves hypothalamic releasing/inhibiting factors. hGH acts on liver and tissues, partly via insulin-like growth factor 1 (IGF-1). Pituitary anatomy distinguishes anterior and posterior lobes with different hormones. Concept / Approach: hGH is produced by somatotroph cells in the anterior pituitary gland. The hypothalamus regulates its secretion through growth hormone–releasing hormone (GHRH) and somatostatin (growth hormone–inhibiting hormone). The pancreas and thyroid produce other hormones (insulin, thyroxine) with separate roles. Step-by-Step Solution: Identify hormone: hGH (somatotropin).Locate source: somatotrophs in anterior pituitary.Note regulation: GHRH stimulates, somatostatin inhibits.Thus, the correct source is the pituitary (anterior lobe). Verification / Alternative check: Clinical testing (GH stimulation/suppression tests) targets pituitary output; pituitary adenomas drive excess GH in acromegaly, confirming the gland as the source. Why Other Options Are Wrong: Hypothalamus: regulates GH but does not secrete the hormone itself into systemic circulation. Pancreas/thyroid: endocrine organs for other hormones. “None of these” is incorrect; the pituitary is correct. Common Pitfalls: Confusing hypothalamic regulation with hormone production; the hypothalamus secretes GHRH and somatostatin, not GH. Final Answer: PITUITARY GLAND (anterior pituitary).

Question 9

Recombinant HBV vaccine platform — The genes encoding HBsAg for the Hepatitis B vaccine were cloned and expressed using which type of vector/host system to mass-produce the antigen?

Accepted Answer

Introduction / Context: The landmark Hepatitis B vaccine leverages recombinant expression of HBsAg in non-pathogenic hosts. Identifying the expression system clarifies why large-scale, safe production was feasible and consistent across global manufacturers. Given Data / Assumptions: HBsAg forms self-assembling virus-like particles when expressed in certain systems. Yeast provides eukaryotic protein processing and scalable fermentation. Vector choice impacts copy number, stability, and yields. Concept / Approach: Many licensed HBV vaccines express HBsAg in Saccharomyces cerevisiae using autonomously replicating yeast plasmids (often 2µ origin-based) under strong promoters. Yeast expression yields high titers of properly assembled HBsAg particles suitable for purification and formulation with alum adjuvant. Step-by-Step Solution: Clone HBsAg gene into yeast expression vector (2µ-based).Transform S. cerevisiae; select high-expressing clones.Ferment, harvest, and purify HBsAg particles.Formulate vaccine; confirm antigenicity and safety. Verification / Alternative check: Product monographs and WHO prequalification summaries describe yeast-derived HBsAg production platforms for multiple licensed vaccines. Why Other Options Are Wrong: General bacterial plasmids: early attempts were less successful for VLP assembly compared to yeast. Cosmids/phagemids: not standard for licensed HBsAg vaccine production. Baculovirus/insect cells: used for some VLPs and proteins, but the classic HBV vaccines are yeast-based. Common Pitfalls: Assuming all recombinant vaccines use bacterial expression; eukaryotic hosts often yield better assembly and post-translational features. Final Answer: Autonomously replicating plasmid of yeast (e.g., 2µ-based; expression in Saccharomyces cerevisiae).

Question 10

Recombinant insulin design — Which approach correctly describes a classic method for synthesizing human insulin using recombinant DNA technology?

Accepted Answer

Introduction / Context: Recombinant human insulin was produced using early bacterial expression strategies that overcame challenges of peptide stability and processing. Understanding the chain-by-chain method explains how bioidentical insulin was obtained without relying on animal pancreases. Given Data / Assumptions: Insulin comprises A and B polypeptide chains linked by disulfide bonds. Bacteria do not process preproinsulin like human beta cells. Fusion expression and chemical cleavage enabled high yields and purity. Concept / Approach: One classic strategy used two synthetic genes (for A and B chains). Each chain was expressed in E. coli as a β-galactosidase fusion, cleaved with cyanogen bromide, purified, and then oxidatively refolded together to form native disulfide bonds. Alternative approved methods later used proinsulin cDNA with enzymatic processing, but the chain-by-chain method is historically significant and matches the option given. Step-by-Step Solution: Chemically synthesize DNA for A and B chains.Express each as a stable fusion in E. coli.Cleave fusions (e.g., CNBr at Met), purify chains.Reconstitute A+B with correct disulfide formation to yield insulin. Verification / Alternative check: Regulatory and historical accounts of Humulin describe chain-by-chain and proinsulin strategies; both yield bioidentical insulin but the option specifying separate chain synthesis matches the classic approach. Why Other Options Are Wrong: Genomic DNA from islets: includes introns and regulatory sequences unsuitable for direct bacterial expression. cDNA without processing: proinsulin requires proper cleavage; expression systems must include processing steps. Single synthetic gene with no processing: still requires folding/processing; oversimplified. Plasma extraction: not feasible for insulin. Common Pitfalls: Assuming bacteria can process preprohormones like human cells; engineered strategies are required to obtain mature insulin. Final Answer: Using chemically synthesized DNA sequences for the A and B chains separately (expressed as fusion proteins and then recombined).

Question 11

Product nomenclature — Human insulin produced by recombinant DNA technology is commonly marketed under which generic name?

Accepted Answer

Introduction / Context: Brand and generic naming conventions help students recognize widely used biopharmaceuticals. Recombinant human insulin revolutionized diabetes care, and its early trade name is still often used as a shorthand in textbooks and examinations. Given Data / Assumptions: “Humulin” refers to human insulin made via recombinant DNA technology. Animal-sourced insulins were previously standard but are now less common. Multiple brands exist, but Humulin is historically emblematic. Concept / Approach: Humulin denotes bioidentical human insulin produced in microbial hosts (E. coli or yeast). It replaced bovine/porcine insulin, improving immunogenicity profiles and supply reliability. Variants (regular, NPH, premixes) share the underlying recombinant origin. Step-by-Step Solution: Identify the rDNA product: human insulin.Recall the historical marketed name: Humulin.Match to the options provided.Select “Humulin.” Verification / Alternative check: Pharmacology texts and formularies list Humulin R and Humulin N as standard preparations of recombinant human insulin. Why Other Options Are Wrong: H insulin/R insulin/huinsulin: not standard trade names. Insugen-RNA: fabricated in this context; not a conventional product name. Common Pitfalls: Confusing “human insulin” (a molecule) with specific analogs (insulin lispro, aspart, glargine) that are engineered variants, not the original Humulin. Final Answer: Humulin.

Question 12

Molecular cloning factoid — In classic vaccine research, the gene coding for the VP1 capsid protein (a major antigenic protein in picornaviruses such as FMDV or poliovirus) was commonly cloned in which standard plasmid vector for expression/analysis?

Accepted Answer

Introduction / Context: VP1 is a key capsid protein in several picornaviruses and an important antigenic target. Early molecular cloning studies frequently used widely available plasmids. Recognizing these classical vectors helps students interpret historic protocols and exam-style questions. Given Data / Assumptions: pBR322 is a pioneering, medium-copy-number plasmid vector with ampicillin and tetracycline resistance markers. pUC series (pUC18/19) are later high-copy derivatives. pMB9 is an early plasmid with limited use compared with pBR322 for teaching examples. Concept / Approach: Many textbook problems and early studies reference pBR322 as the “standard” cloning vector used to insert viral genes like VP1 for subcloning, mapping, and expression in E. coli. While other vectors are certainly possible, pBR322 is the canonical answer in such classic MCQs about VP1 gene cloning from picornaviruses. Step-by-Step Solution: Identify historical context: early cloning era favored pBR322.Associate VP1 cloning with common vectors of the time.Among the listed options, pBR322 is the most canonical choice.Select pBR322. Verification / Alternative check: Older literature and academic exercises often feature pBR322 for inserting viral capsid genes due to its availability and well-characterized restriction map. Why Other Options Are Wrong: pMB9: historically important but less commonly cited in teaching for VP1 cloning. pUC18/pUC19: widely used later; the question’s classic framing usually expects pBR322. pBluescript: versatile but from a later generation and not the stereotypical “first-choice” historical vector. Common Pitfalls: Assuming the newest vector must be correct; exam questions often seek the historically famous pBR322 answer. Final Answer: pBR322.

Question 13

Virology (Picornaviridae) — Within the picornavirus capsid (VP1, VP2, VP3, VP4), which structural protein is the major immunogenic component that elicits neutralizing antibodies (for example, in foot-and-mouth disease virus, the G–H loop)?

Accepted Answer

Introduction / Context: Picornaviruses (such as foot-and-mouth disease virus, poliovirus, rhinoviruses) have icosahedral capsids made of four structural proteins: VP1, VP2, VP3, and VP4. Among these, VP1 is surface-exposed and carries key antigenic sites. Immunology and vaccinology questions often test recognition of which capsid protein induces neutralizing antibodies, because this underpins epitope-focused vaccine design and diagnostic serology. Given Data / Assumptions: Picornavirus capsid is an icosahedral shell with repeating VP1–VP3 on the surface and VP4 on the inner face. Neutralizing epitopes must be accessible to antibodies. Foot-and-mouth disease virus (FMDV) VP1 contains an immunodominant G–H loop region. Concept / Approach: The most immunogenic and neutralization-relevant region must be solvent-exposed. VP1 is prominently displayed on the virion exterior. The famous G–H loop in VP1 (containing an RGD motif in FMDV serotypes) is a classic neutralization epitope. Therefore, VP1 is the best single choice for “immunogenic in nature.” Step-by-Step Solution: Identify which proteins are on the virion surface → VP1, VP2, VP3.Check which harbors dominant neutralizing epitopes → VP1 (G–H loop in FMDV).Conclude that VP1 is the principal immunogenic capsid protein driving neutralizing antibody responses. Verification / Alternative check: Peptide vaccines and recombinant subunits targeting VP1 epitopes of FMDV elicit neutralizing antibodies; escape mutations frequently map to VP1 loop regions, confirming immunological pressure at VP1. Why Other Options Are Wrong: VP2/VP3: surface-exposed but typically less dominant for neutralization than VP1 in picornaviruses.VP4: largely internal on the capsid inner face; not a prime neutralization target.VP0: a maturation precursor that cleaves to VP2 + VP4; not present as such in the mature virion. Common Pitfalls: Assuming any surface protein is equally immunogenic. Dominance depends on epitope exposure and flexibility; in picornaviruses, VP1 generally dominates neutralization. Final Answer: VP1.

Question 14

Foot-and-mouth disease virus (FMDV) genome size — Approximately how many nucleotides are present in the single-stranded RNA (ssRNA) molecule of this picornavirus?

Accepted Answer

Introduction / Context: Picornaviruses possess positive-sense, single-stranded RNA genomes that function directly as mRNA upon entry. Knowing typical genome sizes helps in designing RT-PCR assays, sequencing strategies, and understanding coding capacity (single large polyprotein strategy). Foot-and-mouth disease virus (FMDV) is a classic member of this family affecting cloven-hoofed animals. Given Data / Assumptions: The genome is single-stranded, positive-sense RNA. It encodes one large polyprotein that is post-translationally cleaved. Typical picornavirus genomes fall in the ~7,000–9,000 nucleotide range. Concept / Approach: Recall the canonical size range and select the closest representative value. FMDV genomes are about 8 to 8.5 kilobases in length. Therefore, the best rounded option among those provided is 8,000 nucleotides. Step-by-Step Solution: Identify viral family → Picornaviridae (FMDV).Recall genome form → +ssRNA, polyadenylated, covalently linked VPg at 5’ end.Match to size range → approximately 8 kb. Verification / Alternative check: Reference genomes of various FMDV serotypes consistently report ~8.2–8.5 kb, validating that 8,000 is the closest rounded choice here. Why Other Options Are Wrong: 1,000 or 5,000: too small to encode the polyprotein and regulatory elements.10,000 or 15,000: exceed typical picornaviral genome length; 15,000 approaches paramyxovirus sizes (−ssRNA), not picornaviruses. Common Pitfalls: Confusing “kb” (kilobases) with “kDa” (kilodaltons) or mixing up DNA vs RNA viruses with larger genomes. Final Answer: 8,000.

Question 15

Protein chemistry — During the reconstitution of native insulin from its separate A and B polypeptide chains, which reagent pair is used in sulfitolysis/renaturation workflows to enable correct disulfide bond formation between the chains?

Accepted Answer

Introduction / Context: Insulin consists of two chains (A and B) linked by two inter-chain disulfide bonds and one intra-chain disulfide in the A chain. Classic chemical synthesis and some recombinant workflows generate individual chains that must be recombined to form the correct native disulfide pattern. Sulfitolysis chemistry is widely used to protect cysteines as S-sulfonates and then allow controlled reformation of disulfide bonds upon renaturation. Given Data / Assumptions: Separate A and B chains require correct pairing via disulfide bonds for biological activity. Sulfitolysis creates S–SO3− (S-sulfonate) derivatives to prevent incorrect disulfide scrambling. Reagents such as sodium sulphite and related disulfonate/tetrathionate partners enable reversible protection and subsequent oxidative refolding. Concept / Approach: Identify the reagent pair associated with sulfitolysis and disulfide reformation, rather than inert salts. Sodium sulphite participates in adding sulfonate groups to cysteine thiols; sodium disulphonate (tetrathionate-like partner) drives the reversible modification and later assists in oxidative coupling to re-form native disulfide architecture during chain combination. Step-by-Step Solution: Prepare chain S-sulfonates using sulphite/disulphonate chemistry.Mix A- and B-chain derivatives under controlled pH to promote correct pairing.Oxidatively remove sulfonates to yield native inter-chain and intra-chain disulfides. Verification / Alternative check: Historical insulin reconstitution protocols employ sulphite/disulfonate systems (analogous to sulphite/tetrathionate) to minimize mispairing and maximize yield of bioactive insulin. Why Other Options Are Wrong: b,c,d) Sulphate salts are largely inert in forming the necessary mixed disulfide/sulfonate intermediates.e) Acid and NaCl do not support controlled disulfide chemistry for insulin folding. Common Pitfalls: Confusing disulfide bond formation reagents with denaturants or simple salts; proper redox buffering is essential for correct pairing. Final Answer: Sodium disulphonate and sodium sulphite.

Question 16

Recombinant protein detail — Compared with certain early bacterial preparations, recombinant human growth hormone (hGH) used clinically today typically lacks which terminal residue present in older methionyl-hGH formulations?

Accepted Answer

Introduction / Context: Human growth hormone (hGH) is a 191–amino acid polypeptide. Early recombinant products expressed in E. coli were sometimes produced as methionyl-hGH (somatrem), which contains an extra N-terminal methionine compared with pituitary-derived hormone. Modern recombinant hGH (somatropin) is engineered to match the native 191–amino acid sequence and therefore lacks the terminal methionine addition. Given Data / Assumptions: Pituitary hGH begins with phenylalanine as the first residue in the mature 191–aa sequence. Some early bacterial expression systems initiated translation with an extra N-terminal methionine (192 aa total). Current therapeutics are designed to be structurally identical to endogenous hGH. Concept / Approach: The question contrasts older methionyl-hGH with present-day recombinant hGH that is sequence-identical to human pituitary hormone. Therefore, the key residue “missing” in modern recombinant hGH (relative to methionyl-hGH) is the N-terminal methionine. Choosing “terminal methionine” captures this distinction. Step-by-Step Solution: Recall native hGH length = 191 aa with no extra N-terminal Met.Early somatrem = methionyl-hGH (added Met at N-terminus).Modern somatropin matches native sequence → no extra terminal Met. Verification / Alternative check: Drug labels and protein databases list somatropin as identical to human GH without an N-terminal Met; somatrem historically contained Met and is less commonly used. Why Other Options Are Wrong: Glutamine, lysine, arginine, leucine are not the historical extra residue in early recombinant hGH preparations. Common Pitfalls: Confusing pituitary GH with methionyl-hGH; always check whether a product is somatropin (191 aa) or somatrem (192 aa with Met). Final Answer: Terminal methionine.

Synthesis of Therapeutic Agents Questions