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Role of ComEA in DNA uptake during transformation of competent Bacillus subtilis

Paper ID Volume ID Publish Year Pages File Format Full-Text
21199 43211 2012 5 PDF Available
Title
Role of ComEA in DNA uptake during transformation of competent Bacillus subtilis
Abstract

The role of the competence protein ComEA in DNA uptake during transformation of competent Bacillus subtilis was analyzed by lysed-protoplast transformation (LP transformation). A comEA deletion mutant was constructed by a fusion polymerase chain reaction. Transformants of the mutant were obtained by LP transformation at a frequency of 1.1 × 102 transformants per μg DNA, representing a low relative efficiency of transformation [RET (mutant/wild type)] of 2.7 × 10− 6. This implied an important role of the protein during DNA uptake. When analyzing LP transformation of comEA with a plasmid (5.7 kb), a similar RET (mutant/wild type) of 5.6 × 10− 5 was obtained. Following addition of DNA into the comEA mutant culture, the number of transformants increased at a rate of 0.5 transformants/min, which was very low compared with the wild-type (6.9 × 104 transformants/min). However, even in the comEA mutant, DNA uptake began immediately after addition of DNA. Using co-transformation analysis of the comEA mutant, short linkages at distances of 2–156 kb could be detected, but not long linkages at distances of 671–1662 kb. Taken together, the results indicate that ComEA plays an important role in the transfer of transforming DNA into the DNA channel and in controlling the rate of DNA uptake.

Keywords
Bacillus subtilis; ComEA; DNA uptake; Competent transformation; Uptake velocity; Competence; Linkage analysis; Co-transformation; Kinetic analysis
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Role of ComEA in DNA uptake during transformation of competent Bacillus subtilis
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 113, Issue 6, June 2012, Pages 689–693
Authors
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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
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Full-text PDF Download
Online Support
Any Questions? feel free to contact us