羊源类鼻疽伯克霍尔德菌BPSS1512基因克隆、原核表达及其蛋白的生物信息学分析

doi:10.16431/j.cnki.1671-7236.2017.08.003

Abstract

Abstract:

The experiment was aimed to study the clone and prokaryotic expression of BPSS1512 gene in goat Burkholderia pseudomallei and analyzed its proteins by bioinformatics. The geneome of Burkholderia pseudomallei was used as the template,and the primers were designed by DNAMAN software referring to genomic DNA sequence of Burkholoderia pseudomallei K96243 strain in GenBank (NC_006351.1).The BPSS1512 gene was amplified by PCR and the recombinant plasmid was constructed. Then the expressed protein was analyzed by SDS-PAGE and Western blotting, and the amino acid sequence encoded by BPSS1512 gene was analyzed by softwares such as DNAMAN.The results showed that the BPSS1512 gene was successfully cloned with the length of 1 425 bp,and the recombinant plasmid pET-28a-BPSS1512 was constructed. The optimum conditions for induction was that the IPTG was 10 mmol/L and 8 h for induction.The molecular weight of the protein was 53 ku,it was expressed as the form of inclusion body.In the secondary structure of BPSS15122 protein,alpha-helix,extended strand,and random coil were 24.05%,14.77% and 61.18%, respectively,and the hydrophobic core was distributed between -2.0 and +2.4 which indicated that the BPSS1512 protein was strong hydrophobicity.

Key words: Burkholderia pseudomallei; BPSS1512 gene; clone; prokaryotic expression; bioinformatics analysis

CLC Number:

Q786

CAO Rui-yong, NIE Xin, LI Bao-bao, ZHANG Zhen-xing, HUANG Hai-feng, LI Ya-ying, PENG Dong-mei, LI Guo-hua, ZHU Shu, YANG Xiao-jian, DU Li, WANG Feng-yang. Cloning,Prokaryotic Expression of BPSS1512 Gene in Goat Burkholderia pseudomallei and Bioinformatics Analysis of its Proteins[J]. , 2017, 44(8): 2234-2240.

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Cloning,Prokaryotic Expression of BPSS1512 Gene in Goat Burkholderia pseudomallei and Bioinformatics Analysis of its Proteins

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