fulltext.study @t Gmail

Random strand transfer recombination (RSTR) for homology-independent nucleic acid recombination

Paper ID Volume ID Publish Year Pages File Format Full-Text
25251 43563 2007 11 PDF Available
Title
Random strand transfer recombination (RSTR) for homology-independent nucleic acid recombination
Abstract

Random strand transfer recombination (RSTR) relies on the ability of viral reverse transcriptases to undergo homology-independent template switches during DNA synthesis. To facilitate strand transfer events single stranded template DNA was hybridized via noncoding complementary sequence stretches which flank the genes. The resulting bulb shaped heteroduplex was used as template for the reverse transcriptase driven DNA polymerization in which template switch events occur spontaneously. Switches were promoted by induced pausing of the viral polymerase at digoxygenin labeled nucleotides. RSTR of homologous genes was demonstrated by the recombination of two genes encoding (R)-hydroxynitrilase isoenzymes from Prunus amygdolus. For non-homologous RSTR we recombined the genes of the Havea brasiliensis (S)-hydroxynitrilase and the estC gene from Burkholderia gladioli. Base-pairing dependent recombination took place spontaneously and at high frequency between genes with low and high sequence homology.

Keywords
Homologous recombination; Non-homologous recombination; Protein engineering; Promoter engineering; Strand transfer; Template switch; Directed evolution
First Page Preview
Random strand transfer recombination (RSTR) for homology-independent nucleic acid recombination
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 129, Issue 1, 30 March 2007, Pages 39–49
Authors
, , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us