Recombinant DNA Technology Overview

Recombinant DNA Technology Overview

• Recombinant DNA technology enables the creation of novel DNA molecules for research and therapeutic purposes.
• Key tools include bacterial plasmid vectors, restriction enzymes, and ligases, pioneered by Stanley Cohen and Herbert Boyer in the 1970s.
• Herbert Boyer co-founded Genentech, the first company to commercialize recombinant DNA technology, igniting the biotechnology sector.

Historical Milestones

• In 1978, Genentech achieved a significant milestone by cloning the human insulin gene into a plasmid vector, allowing insulin production in E. coli.
• Prior to this, insulin extraction from cows was demanding and required modifications to prevent immune rejection.
• In 1982, humulin, the first recombinant human insulin, received FDA approval.

Basic Process of Recombinant DNA Creation

• A vector DNA molecule is manipulated in vitro to insert foreign DNA, forming a recombinant plasmid.
• This plasmid can be introduced into bacteria for replication alongside the bacterial DNA.
• Desired proteins like insulin can be isolated, or recombinant plasmids can be analyzed for foreign DNA sequences.

Application in Water Quality Research

• The focus is on sequencing a segment of the 16s ribosomal RNA gene to identify microbial diversity in the Sacramento River.
• The Sacramento River, California's largest watershed, faces contamination from urban runoff and agriculture, affecting water quality.
• Understanding microbial populations is vital as certain species can indicate ecological health and contribute to detoxifying pollutants.

Project Role and Cloning Definition

• The project involves cloning a 16s rRNA gene fragment previously amplified via PCR by biology students.
• Cloning here refers to inserting a PCR fragment into a vector, leading to recovery of the recombinant plasmid from a bacterial colony.