A SOE-PCR method of introducing multiple mutations into Mycoplasma gallisepticum neuraminidase

Lei Tan; Hongjun Chen; Shengqing Yu; Xusheng Qiu; Cuiping Song; Danqing Chen; Shilei Zhang; Fanqing Zhang; Suibin He; Xinyue Shen; Meirong Hu; Chan Ding

doi:10.1016/j.mimet.2013.05.013

A SOE-PCR method of introducing multiple mutations into Mycoplasma gallisepticum neuraminidase

J Microbiol Methods. 2013 Aug;94(2):117-120. doi: 10.1016/j.mimet.2013.05.013. Epub 2013 May 24.

Authors

Affiliations

¹ Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, P. R. China.
² Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, P. R. China; Poultry Institute, Chinese Academy of Agricultural Sciences, Yangzhou 225009, P. R. China.
³ Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, P. R. China. Electronic address: shoveldeen@shvri.ac.cn.

PMID: 23707236
DOI: 10.1016/j.mimet.2013.05.013

Abstract

A modified splicing with overlap extension PCR (SOE-PCR) was generated to introduce 21 TGA to TGG at Mycoplasma gallisepticum MGA_0329 gene. The recombinant protein was successfully expressed and retained neuraminidase activities, indicating that SOE-PCR is a rapid and highly efficient method of introducing multiple mutations into large M. gallisepticum genes.

Keywords: M. gallisepticum; MGA_0329 gene; Neuraminidase; SOE-PCR.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Proteins / genetics*
Bacterial Proteins / metabolism
Mutagenesis, Site-Directed / methods*
Mycoplasma gallisepticum / enzymology*
Mycoplasma gallisepticum / genetics
Neuraminidase / genetics*
Neuraminidase / metabolism
Point Mutation*
Polymerase Chain Reaction / methods*

Substances

Bacterial Proteins
Neuraminidase