猪附红细胞体ENO基因重组腺病毒穿梭质粒的构建及真核表达

doi:10.16431/j.cnki.1671-7236.2018.08.009

Abstract

Abstract:

To construct recombinant adenovirus shuttle plasmid of ENO gene of Mycoplasma suis and express it in eukaryotic cells,a pair of primers was designed and synthesized according to the ENO gene sequence of Mycoplasma suis in GenBank (accession No.:CP002525.1) in this study.ENO gene was amplified by PCR method,and the products were cloned into the pMD19-T vector.The pMD19T-ENO plasmid was digested with Kpn Ⅰ and Xho Ⅰ, and then subcloned into the adenovirus shuttle vector PCR259.The PCR259-ENO plasmid was constructed and then identified by PCR and enzyme digestion.The PCR259-ENO recombinant shuttle plasmid was transfected into 293 cells by liposome-mediated transfection,and the expression of ENO gene in 293 cells was detected by IFTA technology.The results showed that the cloned ENO gene was 1 182 bp in length and encoded 393 amino acids with a homology of 99% with the ENO gene sequence in GenBank (accession No.:CP002525.1).The constructed recombinant adenovirus shuttle plasmid PCR259-ENO was identified by PCR and restriction enzyme digestion correctly, and was able to be expressed in 293 cells,indicating that ENO gene was successfully inserted into adenovirus shuttle plasmid PCR259 and the recombinant shuttle plasmid was successfully constructed.

Key words: Mycoplasma suis; ENO gene; recombinant adenovirus shuttle plasmid; expression

CLC Number:

Q784

WANG Miao, NI Tingting, WU Shengjun, ZHAO Yun, SONG Jianchen, DONG Liang, XUE Shujiang. Construction and Eukaryotic Expression of Recombinant Adenovirus Shuttle Plasmid of Mycoplasma suis ENO Gene[J]. , 2018, 45(8): 2113-2118.

References

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Construction and Eukaryotic Expression of Recombinant Adenovirus Shuttle Plasmid of Mycoplasma suis ENO Gene

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