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Construction and Expression Analysis of Prokaryotic Expression Vector of UPA3-0382 Gene
He Conghui1,3, Li Chunliang2, Li Xueyan1,3, Jia Zewei1,3, Li Xiaoyan1,3, Jia Tianjun1,3*
1College of Medical Laboratory, Hebei North University, Zhangjiakou 075000, China; 2Hebei Northern University Affiliated Hospital, Zhangjiakou 075000, China; 3Institute of Pathogenic Biology and Immunology, Hebei North University, Zhangjiakou 075000, China
Abstract: In order to investigate the function of Ureaplasma urealyticum UPA3-0382 gene， a prokaryotic expression vector of Ureaplasma urealyticum UPA3-0382 was constructed and bioinformatics analysis of the expressed protein was performed in this study. At first， UPA3-0382 gene with a length of 624 bp was cloned by PCR， the target gene and plasmid pGEX-6P-2 were digested by BamH I and Not I， then recombined with T4 ligase， the recombinant plasmid was transformed into XL1-Blue and then expressed fusion-protein. Finally， the structure and function of the fusion protein was predicted with bioinformatical software. The results showed that the homology between sequencing results and UPA3-0382 in NCBI was 100% and the molecular weight of fusion protein was 50 kDa. UPA3-0382 gene encoding protein consists of 207 amino acids and its molecular formula was C1075H1749N305O348S3， the molecular mass was 24.62 kDa and its theoretical isoelectric point was 6.40. The ratio of α-helix， β-fold， β-turn and random curl in the secondary structure of the protein was 48.79%， 16.91%， 7.25% and 27.05%， respectively. There is no transmembrane domain and no signal peptide in this protein. Protein spatial structure and function predictions show that this protein is a hypothetical protein and may participate in Ureaplasma urealyticum signal transduction and energy metabolism. These fingdings may lay the foundation for further study of Ureaplasma urealyticum.
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