鹌鹑VIPR-1的克隆、序列特征和组织表达分析

doi:10.3864/j.issn.0578-1752.2012.03.015

Abstract

Abstract: 【Objective】Vasoactive intestinal peptide type 1 receptor (VIPR-1) plays an important role in poultry reproduction, but the gene and its analysis have not been reported in quail prior to this study. The objective of this study was to clone the cDNA of quail VIPR-1, analyze its sequence and tissue expression pattern. 【Method】 Based on comparative genomics, the cDNA sequence of quail VIPR-1 was obtained by RT-PCR and RACE PCR. The nucleotide and amino acid sequences were analyzed and compared with that of other species. The expression levels of VIPR-1 in eight quail tissues were detected by real-time quantitative PCR. 【Result】 A 2 427 bp cDNA of the quail VIPR-1 was cloned and the length of its coding region was 1 341 bp, encoding a receptor precursor of 446 amino acids. There are 41 nucleotide differences in the coding region of VIPR-1 between quail and chicken, causing 4 amino acid differences in VIPR-1 protein. The quail VIPR-1 shows high amino acid sequence identity to that of chicken (99.1%), turkey (92.2%), and zebra finch (88.0%). Meanwhile, quail VIPR-1 also shares 60%-78% amino acid sequence identity to that of other species. The phylogenetic tree of VIPR-1 was consistent with the evolutionary relationship among species. The physical and chemical properties of quail VIPR-1 indicated that VIPR-1 was a basic protein. The secondary structure of quail VIPR-1 was made of α-helixes, β-sheets and β-turns. This protein contains a putative signal peptide (MKSARLRVLLPLLGCLLSAASS) in its N-terminus, 7 transmembrane domains, and a C-terminal hydrophilic domain. Real-time quantitative PCR assay demonstrated that the VIPR-1 mRNA could be detected in all 8 quail tissues examined with the highest expression level being noted in small intestinal.【Conclusion】The full length cDNA of quail VIPR-1 was cloned and the highest expression level of VIPR-1 was detected in small intestinal. In addition, the cholesyerol binding sites were identified on transmembrane domains of quail VIPR-1.

Key words: quail, VIPR-1, molecular characteristics, tissue expression

ZHOU Min, LI Ying, SHEN Xu, XU Hai-Ping, ZHANG Cheng-Guang, ZHANG Xi-Quan. cDNA Cloning, Sequence Analysis and Tissue Specific Expression of Vasoactive Intestinal Peptide Type 1 Receptor (VIPR-1) in Quails[J].Scientia Agricultura Sinica, 2012, 45(3): 529-539.

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cDNA Cloning, Sequence Analysis and Tissue Specific Expression of Vasoactive Intestinal Peptide Type 1 Receptor (VIPR-1) in Quails

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