@article{ref1,
title="Construction of hemolysin hlyA gene knocked out of cholera vaccine candidate and inserted green fluorescent protein gene",
journal="The Journal of Practical Medicine",
year="2016",
author="Zhou, Yanyan and Kan, Biao and Rui, Yongyu",
volume="",
number="24",
pages="362-366",
abstract="OBJECTIVE To knock down hemolysin hlyA gene and insert green fluorescence protein gene of vibrio cholerae vaccine candidate NFYY101. <br><br>METHODS Amplified fragments of hlyA gene upstream (hlyAup) and downstream (hlyAdown),lacz-GFPuv,and hlyAup-laczGFPuv-hlyAdown, and plasmids treated with specific enzymes were utilized to construct recombinant plasmids pUC18-hlyAup-laczGFPuv-hlyAdown and pCVD442-hlyAup-laczGFPuv-hlyAdown. Following the construction of the recombinant suicide plasmids ,a vaccine candidate was constructed by homologous recombination ,while SM10λpir carrying pCVD442- hlyAup-laczGFPuv-hlyAdown was utilized as the donor strain to transfect NFYY101. <br><br>RESULTS Construction of recombinant suicide plasmids pCVD442- hlyAup-laczGFPuv-hlyAdown was satisfactory that hemolysin hlyA gene was knocked out and green fluorescence protein gene was successfully inserted of vibrio cholerae vaccine candidate NFYY101. <br><br>CONCLUSION Construction of the recombinant suicide plasmid pCVD442-hlyAup-laczGFPuv-hlyAdown successfully can be used for knocked out the hlyA gene and added green fluorescence protein gene as genetic marker into the chromosomal DNA of vibrio cholerae vaccine candidate.<p /><p>Language: zh</p>",
language="zh",
issn="1006-5725",
doi="10.3969/j.issn.1006-5725.2016.03.007",
url="http://dx.doi.org/10.3969/j.issn.1006-5725.2016.03.007"
}