Construction of Soybean Peroxidase Gene Random Mutation Library(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:

2011年05期

Page:

731-737

Research Field:

Publishing date:

Info

Title:

Construction of Soybean Peroxidase Gene Random Mutation Library

Author(s):

ZENG Jia-yu¹; ZHENG Qun¹; LIAO Shi-qi²; WANG Li²; YUAN Hong-xia²; YANG Hai-tang¹; ZHANG Li-qiong³; ZHANG Ning³

1.College of Life Science，Northwest Normal University，Lanzhou 730070;?

2.Gansu Province Institute of Medical Sciences，Lanzhou 730050；

3.School of Life Science and Engineering，Lanzhou University of Technology，Lanzhou 730050，Gansu，China

Keywords:

Soybean peroxidase; Error-prone PCR; Gene library; Random mutation; Directed evolution of enzymes

PACS:

S565.1

DOI:

10.11861/j.issn.1000-9841.2011.05.0731

Abstract:

In order to improve enzymatic activity of soybean peroxidase(SBP)and study the correlation between its primary structure and enzymatic activity，sequential error-prone PCR method was used on soybean peroxidase genes（sbp） for two times randon mutation.The second round error-prone PCR products and plasmid vectors pPICZα-A were digested by double enzymes.After the PCR products ligated into the vectors by T₄DNA ligase，recombinant plasmids pPICZα-A-sbp were transformed into Escherichia coli DH5α through calcium chlorid method.A random mutation library was constructed，and the capacity was about 1.77×106 cfu.

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