水牛甲状旁腺素1受体基因CDS区克隆与生物信息学分析

doi:10.16431/j.cnki.1671-7236.2018.02.002

Abstract

Abstract:

In order to enrich the basic data of buffalo parathyroid hormone 1 receptor (PTH1R) gene, the CDS region of buffalo PTH1R was cloned, and the sequence was analyzed by bioinformatics method in this study. Primers sequence were designed according to cDNA sequence of cattle PTH1R gene in GenBank (accession No:NM_001075332.1) by Primer Premier 5.0 software. The CDS region in buffalo PTH1R gene was cloned by PCR amplification and nucleic acid sequencing technology. The primary structure, secondary structure, tertiary structure, physicochemical properties, homology were analyzed and phylogenetic tree of PTH1R was constructed by DNAMAN, ProtParam, SOPMA and PSORTⅡ prediction softwares. The results showed that the length of PTH1R gene in buffalo was 2 283 bp, the CDS region was 1 770 bp, GenBank accession No.was MF380401, encoding 589 amino acids. The homology of buffalo PTH1R gene CDS region compared to cattle, pigs, horses, goats, sheep and camels were 99.4%, 93.2%, 93.5%, 95.3%, 98.1% and 93.9%, respectively. There was a high homology among different species, and phyletic evolution was in accordance with their genetic relationship. The research indicated that PTH1R gene was conservative in the course of evolution. Analysis of physicochemical properties of protein showed that PTH1R protein molecular formula was C₂₉₉₆H₄₆₁₆N₇₉₂O₈₂₃S₃₀, the molecular weight was 65 860.22 u, the half-life was 30 h, theoretical electrical points (pI) was 8.37, the extinction coefficient of water solution at 280 nm was 117 770, the peptide chain N-terminal was M (Met), the unstable coefficient was 43.36, belong to the basic unstable protein. The fat coefficient of PTH1R protein was 88.61, and the total average hydrophilicity was 0.007, belongs to insoluble protein. The secondary structure analysis showed that the buffalo PTH1R gene protein contained 218 α-helixes, 114 extended strands, 44 β-turns and 213 random colis, and consistent with the tertiary structure prediction. PTH1R gene coding region was conservative in the course of long-term biological evolution. This results provided a theoretical basis for revealing the genetic characteristics of buffalo PTH1R gene.

Key words: buffalo; PTH1R gene; cloning; bioinformatics analysis

CLC Number:

LU Xingrong, LIANG Xianwei, LIANG Shasha, DENG Tingxian, DUAN Anqin, MA Xiaoya, PANG Chunying. Cloning and Bioinformatics Analysis on PTH1R gene CDS in Buffalo[J]. , 2018, 45(2): 291-301.

References

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Cloning and Bioinformatics Analysis on PTH1R gene CDS in Buffalo

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