Chat with Har Gobind Khorana

Biochemist and Nobel Laureate

About Har Gobind Khorana

In a cramped University of Wisconsin lab in the late 1960s, with hand-poured acrylamide gels and radioactive phosphorus painstakingly tracked on X-ray film, he orchestrated the chemical synthesis of a functional gene—69 nucleotides long—encoding the yeast alanine tRNA. This wasn’t theoretical; it was tactile, iterative biochemistry: cutting DNA with restriction enzymes before they were commercially available, ligating fragments with polynucleotide kinase he purified himself, verifying sequence fidelity through partial digestion and two-dimensional electrophoresis. His codon assignments weren’t inferred from statistical correlations alone—they emerged from triplet-binding assays using ribosomes, synthetic RNA polymers like poly-U and poly-UC, and radiolabeled aminoacyl-tRNAs, each experiment calibrated against known protein sequences. He viewed nucleic acids not as abstract code but as chemically addressable polymers—each phosphate bond a handle, each base pair a stoichiometric partner. That mindset bridged enzymology and information theory, turning genetic decipherment into an engineering discipline long before CRISPR or next-gen sequencing existed.

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Conversation Starters

Not sure where to begin? Try asking Har Gobind Khorana:

  • “How did you confirm UUU coded for phenylalanine without modern sequencing?”
  • “What made you choose tRNA over mRNA for your first synthetic gene?”
  • “Why did you insist on purifying your own polynucleotide kinase in 1967?”
  • “Did the 1968 Nobel Prize change how you approached collaborative science?”

Frequently Asked Questions

Did Khorana’s team synthesize the entire yeast alanine tRNA gene in one piece?
No—the gene was assembled in overlapping segments: three 20–25 nucleotide fragments synthesized chemically, then ligated stepwise using T4 DNA ligase and kinase. The final 77-nucleotide construct included promoter elements and was cloned into a plasmid only after verification by nuclease digestion mapping and hybridization. Full-length synthesis wasn’t feasible until solid-phase methods matured in the 1980s.
What role did Khorana play in developing the first DNA synthesizer?
He didn’t build or commercialize a synthesizer—but his 1970 PNAS paper established the critical phosphotriester method for stepwise oligonucleotide assembly, directly informing Marvin Caruthers’ later phosphoramidite chemistry. Khorana’s group optimized protecting groups (e.g., dimethoxytrityl) and coupling reagents under anhydrous conditions, solving stability issues that had stalled prior efforts.
Why did Khorana shift from codon work to tRNA synthesis in the early 1970s?
He recognized that codon assignments meant little without understanding how tRNAs physically decoded them. Synthesizing tRNA allowed him to test anticodon mutations, study conformational switching upon aminoacylation, and probe ribosome binding kinetics—transforming the genetic code from a static table into a dynamic molecular mechanism.
How did Khorana’s upbringing in pre-partition India influence his experimental style?
Trained in Lahore’s Government College where lab equipment was scarce, he learned to improvise: distilling solvents over charcoal, calibrating centrifuges with stopwatch-and-string timing, repurposing silk-screen mesh for gel filtration. That resourcefulness persisted—he built custom chromatography columns from glass tubing and silica gel he activated in his lab oven, prioritizing functional precision over commercial instrumentation.
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Topics

biochemistrygenetic codenobel prizemolecular biologyscience history

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