郏县红牛POLB基因克隆及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2022.03.010

Abstract

Abstract: 【Objective】 The aim of this study was to clone Jiaxian Red cattle DNA polymerase beta (POLB) gene and analyze its bioinformatics.【Method】 The complete CDS region of cattle POLB gene was cloned by PCR with the cDNA of muscle tissue in Jiaxian Red cattle as the template.Genetic distance analysis and phylogenetic tree were constructed by online softwares to analyze the physical and chemical properties, hydrophobicity, phosphorylation sites, transmembrane structure, signal peptide, secondary structure, tertiary structure and interaction proteins of POLB protein.【Result】 The sequence of CDS region of Jiaxian Red cattle POLB gene was 1 008 bp, encoding 335 amino acids.Phylogenetic tree analysis showed that the amino acid sequence of Jiaxian Red cattle POLB gene was closest related to sheep and other ruminants, followed by other mammals and poultry, and was farthest related to zebrafish (fish).The molecular weight of Jiaxian Red cattle POLB protein was 38.26 ku, the theoretical isoelectric point (pI) was 9.10, the half-life was 30 h, the instability coefficient was 31.06, and the total average hydrophilic coefficient was -0.659, indicating that it was a stable hydrophilic protein.Predictive analysis of phosphorylation sites showed that Jiaxian Red cattle POLB protein contained 17 serine phosphorylation, 12 threonine phosphorylation and 4 tyrosine phosphorylation sites.The prediction results of transmembrane region and signal peptide showed that Jiaxian Red cattle POLB protein did not belong to transmembrane protein and secretory protein.The prediction of secondary structure and tertiary structure showed that the structure mainly consisted of alpha helix, beta turn, extended strand and random coil.POLB protein was predicted to interact with XRCC1, FEN1, PARP1 and LIG3 by online softwares.【Conclusion】 The CDS sequence of Jiaxian Red cattle POLB gene was successfully cloned in this study, and it was closest to sheep.POLB protein was a hydrophilic protein without transmembrane structure and signal peptide, and had the highest degree of interaction with XRCC1 protein.The results could provide references for further exploring the biological function of POLB gene.

Key words: Jiaxian Red cattle; POLB gene; cloning; bioinformatics analysis

CLC Number:

LI Yulong, LYU Shijie, ZHAI Yaying, ZHANG Zhijie, ZHU Xiaoting, ZHANG Zijing, CHEN Fuying, SHI Qiaoting, WANG Eryao. Cloning and Bioinformatics Analysis of POLB Gene in Jiaxian Red Cattle[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 887-896.

References

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Cloning and Bioinformatics Analysis of POLB Gene in Jiaxian Red Cattle

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