双峰驼CYP2J cDNA全长克隆、序列信息及组织表达分析

doi:10.11843/j.issn.0366-6964.2018.10.009

Abstract

Abstract:

In order to study the structure and function of CYP2J gene and protein of Bactrian camel, the full-length cDNA sequence of CYP2J gene of Bactrian camel was amplified firstly by RACE and RT-PCR techniques. Then the bioinformatics method was used to analyze the characteristics of the protein encoded by CYP2J gene, and the phylogenetic tree of CYP2J protein between Bactrian camel and related species was constructed by using MEGA 6. Finally, real-time quantitative PCR was used to determine the difference of CYP2J gene expression in various tissues of Bactrian camels. The results showed that the full-length CYP2J cDNA sequence of Bactrian camel was 1 770 bp containing a 1 509 bp complete open reading frame (ORF), which encoded a protein with 502 amino acids. Bioinformatics analysis indicated that the protein encoded by the ORF was predicted to be a stable hydrophilic protein without signal peptide recognition function but had a transmembrane segment (12-35 aa), and was anchored to endoplasmic reticulum. And some important physicochemical properties of the protein such as the molecular formula, isoelectric point, molecular weight, number of positively charged residues (Arg+Lys) and negatively charged residues (Asp+Glu) were C₂₆₄₅H₄₀₉₉N₇₁₃O₇₂₆S₁₅, 8.45, 57 983, 59 and 56, respectively. The secondary structure were mainly composed of 46.41% alpha helix, 13.15% extended strand and 40.44% random coil. Further phylogenetic tree analysis indicated that CYP2J protein of Bactrian camel had the closest genetic relationship with CYP2J protein of sheep and cattle. The real-time quantitative PCR result showed that CYP2J gene expressed in every tissues detected of Bactrian camel and the expression level of CYP2J gene was the highest in liver and pancreas, followed by kidney, lung, heart, small intestine and blood vessels, and the lowest in spleen and large intestine. In conclusion, the full-length cDNA sequence of CYP2J of Bactrian camel and the bioinformation of protein encoded by open reading frame(ORF) were successfully obtained in this study, which laid the foundation for preparing the CYP2J protein and antibody of Bactrian camel.

CLC Number:

S824.2

JIA Zhi-peng, PENG Xiao-bin, LUO Cha-gan, HASI Su-rong. The Full-length Cloning, Sequence Information and Tissues Expression Analysis of CYP2J cDNA in Bactrian Camel[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(10): 2133-2144.

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The Full-length Cloning, Sequence Information and Tissues Expression Analysis of CYP2J cDNA in Bactrian Camel

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