RECOMBINANT 2

9問 • 1年前

問題一覧

Cloning Insulin A & B Chains Using Recombinant DNA Technology 1. Isolation of the Insulin Gene The human insulin gene is extracted from human _______ The gene sequence for the A-chain and B-chain is _________. Alternatively, the genetic code for the A and B chains can be synthesized artificially if the complete human insulin gene is unavailable.

pancreatic cells; identified and isolated

Cloning Insulin A & B Chains Using Recombinant DNA Technology 2. Preparation of Plasmid Vector A ______ (a small, circular DNA found in bacteria) is used as a vector to carry the insulin gene. The plasmid is cut using a _______ at specific sites, creating openings where the A and B chain gene sequences can be inserted.

plasmid; restriction enzyme

Cloning Insulin A & B Chains Using Recombinant DNA Technology 3. Insertion of A & B Chain Genes into Plasmid The isolated genes for the A-chain and B-chain are inserted separately into different plasmid vectors. The enzyme DNA ligase is used to "glue" the _______ into the plasmid. This creates two recombinant plasmids — one carrying the A-chain gene and the other carrying the B-chain gene.

A-chain and B-chain genes

Cloning Insulin A & B Chains Using Recombinant DNA Technology 4. Transformation into Host Bacteria The recombinant plasmids are introduced into ______ (usually Escherichia coli). The bacteria take up the plasmids through a process called ______, becoming "genetically modified" to produce human insulin chains.

host bacteria; transformation

Cloning Insulin A & B Chains Using Recombinant DNA Technology 5. Production of A & B Chains Inside the bacteria, the ______ are expressed, and the A-chain and B-chain proteins are produced separately. The bacteria are cultured in large bioreactors, allowing them to multiply and produce large quantities of the A and B insulin chains.

plasmids

Cloning Insulin A & B Chains Using Recombinant DNA Technology 6. Isolation and Purification of Insulin Chains The A and B chains are extracted from ______. The ______ are purified to remove bacterial proteins, toxins, and contaminants. Each chain is tested to ensure it is correctly folded and functional.

bacterial cells; proteins

Cloning Insulin A & B Chains Using Recombinant DNA Technology 7. Combining A & B Chains to Form Insulin The purified A and B chains are mixed under specific conditions that promote the formation of disulfide bonds between them. These disulfide bonds link the A-chain and B-chain, resulting in the formation of functional _______

human insulin

Cloning Insulin A & B Chains Using Recombinant DNA Technology 8. Final Purification and Quality Control The final insulin product is purified again to ensure it is free from _____. Rigorous quality control tests are conducted to ensure safety, efficacy, and purity. The purified insulin is then packaged and made available for use in treating diabetes.

impurities

This method, pioneered by _______ in 1978, was the first successful commercial production of synthetic human insulin, branded as Humulin. Before this, insulin was extracted from the pancreases of pigs and cows, which often caused allergic reactions in humans. The _________ revolutionized diabetes treatment by providing a purer, safer, and more efficient source of insulin.

Genentech; recombinant DNA method

CH 1

OBILLO, MIKAELA T. · 78問 · 1年前

CH 1