Cloning and Expression of Pediocin Gene

doi:10.7668/hbnxb.2009.02.007

Abstract

Abstract: Plasmid DNA was extracted from Pediococcus acidilactici as template.The oligonucleotide primers were designed according to the gene sequence reported by GenBank.Amplied the DNA fragment by means of PCR and cloned to pTA2 Vector to sequenced.Compared the inserted DNA fraction and the Ped-A sequence reported by GenBank,and found sequence homology was 100%.Then inserted the segment to expressing vectorPet-28a.The recombinant plasmid transformed to Escherichia coli DH5,screened by kanamycin resistance and identified by PCR,restriction endonucleases analyzing,obtained positive recombinant plasmid transformed which to E. coli BL21(DE3) receptivity cell,induced with IPTG,Tricine-SDS-PAGE showed its molecularmass as 4 600 Dathe same as thereported pediocin PA-1.

Key words: Pediococcus acidilactici, PCR, Cloning, Heterogenous expression

CLC Number:

Q939.11+7

HAN Ye, ZHOU Zhi-jiang, WANG Pei-pei, ZHANG Li-xia. Cloning and Expression of Pediocin Gene[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(2): 32-35. doi: 10.7668/hbnxb.2009.02.007.

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