Use of quantitative PCR technique for determining gene copy number in the genome of Beauveria bassiana transformant
Dong, Wei-Xia | Feng, Ming-Guang | Ying, Sheng-Hua
This study describes the use of quantitative PCR (qPCR) to determine gene copy number in Beauveria bassiana genome. This procedure has been improved by generating a reference strain to calibrate PCR amplification efficiency and using a γ-tubulin gene as internal control. The reference strain was constructed by introducing one copy of phosphinothricin resistance gene and two copies of green fluorescent protein gene into the wild-type strain. Finally, this improved procedure was validated via determining the gene copy number in the previously characterized transformants, and displays a great potential in rapid detection of gene copy number in the genome of transformant.
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