TRANSFORMATION PROCEDURE

Asked April 10, 2016, 6:35 PM EDT

Hello, my name is June Hernandez & I am a freshman enrolled in the biotechnology program at my school. Okay, so my question is, what would be the effects on a plasmid/organism if a gene is incorrectly inserted? If the transformation procedure fails, what would happen and how could it be explained? I would like to go ahead and thank you in advance. I understand I have no connection to you, but I would greatly appreciate your help. Thank you for taking time of your day.

Dallas County Texas genetics

1 Response

Hi June, neat question! The typical laboratory methods for inserting genes using vectors like plasmids are inexpensive and fairly simple to do, but don't allow the insertion site to be targeted to one specific site in a genome. Random insertion of these new sequences means that most attempts fail. For example, if you are tying to insert a gene in to an E. coli bacterium most bacteria will not take up the vector, some with take up the vector but the insertion will occur in a dangerous place in the genome and lead to death, and very few bacteria will successfully incorporate the new sequence in to their genome. Attempts to use "gene therapy" to cure heritable diseases have struggled with this limitation and in some experiments random insertion of new, "healthy", genes has led to complications like cancer.

Fortunately, very recently developed methods like "CRISPR" are enabling precisely targeted insertion of genetic sequences. CRISPR is already accelerating research and may soon offer some hope to those suffering from genetic disease.

Good luck in your Biotech program! -SB