›› 2017, Vol. 44 ›› Issue (6): 1604-1612.doi: 10.16431/j.cnki.1671-7236.2017.06.005

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Nested PCR Amplification and Bioinformatics Analysis of LSm1 Gene from Marc145 Cells

CHEN Shao-pin1,2, LIN Han-qing1,2, WEN Gui-lan1,2, ZHANG Sheng-bo1,2, LI Chang-hong1,2, XU Li1,2, GONG Xin-yong3, WANG De-sheng1,2, WEN Ming1,2, ZHOU Bi-jun1,2, CHENG Zhen-tao1,2   

  1. 1. College of Animal Science, Guizhou University, Guiyang 550025, China;
    2. Key Laboratory of Animal Disease and Veterinary Public Health of Guizhou Province, Guiyang 550025, China;
    3. College of Information and Information Engineering, Guizhou University, Guiyang 550025, China
  • Revised:2017-03-27 Online:2017-06-20 Published:2017-06-28

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Abstract:

In order to analyze the character of LSm1 gene and predict its biology function in this study,LSm1 gene from the Marc145 cells was amplified and cloned by nested PCR,and the secondary structure, B cell epitopes,conserved domain, transmembrane domain and signal peptide of LSm1 protein were predicted by bioinformatics softwares. The results showed that LSm1 gene sequence with 402 bp was successfully obtained,which could encode 133 amino acids. The nucleotide sequence of LSm1 gene from Marc145 cells was high similarity with human and primates,followed by marine mammals,terrestrial wild animal and livestock,the percent identity was from 92.8% to 99.8%.The encoding amino acid sequence similarity was from 97.8% to 99.3%. The results of phylogenetic tree showed that the LSm1 gene of Marc145 cells was closely relative with human which were in the same branch,followed by primates. The alpha helix and random coil were dominant in secondary structure ,which was 45.11% and 24.06%,respectively. It was predicted that there were 4 B cell dominant epitopes and a conserved domain of Sm superfamily in the LSm1 protein, and there was no transmembrane region and signal peptide region. In conclusion,the LSm1 protein might has low transcription and expression level in the intracellular,and nested PCR was needed to achieve the target band from cDNA of cells. The method provided a reference for the amplification of LSm1 gene. At the same time,the prediction results of it's biological function would lay a foundation for studying the LSm1 artificial expression protein,the antibody and the functional mechanism of LSm1 at the cellular level.

Key words: LSm1 gene; nested PCR; cloning; bioinformatics analysis

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