天柱番鸭NPY基因克隆及其在组织中的表达分析

doi:10.16431/j.cnki.1671-7236.2019.04.002

Abstract

Abstract:

This study was aimed to clone neuropeptide Y (NPY) gene in Tianzhu muscovy,and analyze its transcriptional level of different tissues in Tianzhu muscovy.The mixed cDNA of Tianzhu muscovy tissues during lying was used as PCR template,then the entire CDS region of NPY gene was cloned and sequenced by RT-PCR.The homology and relationship of genetic evolution of NPY gene,the physical and chemical properties,subcellular localization,signal peptide,glycosylation site and phosphorylation site,the secondary and tertiary structures of NPY protein were analyzed by bioinformatics tools,and the transcriptional level of NPY gene mRNA in different tissues were detected.The results showed that the whole-length coding region of NPY gene in Tianzhu muscovy was 294 bp,which coded 97 amino acids.The homology alignment results of nucleotide and amino acid sequences showed that there was a certain genetic diversity of NPY gene in different species,and Tianzhu muscovy had a close relationship with Anas platyrhynchos.NPY protein were acidic and unstable protein,which had one signal peptide (1-28 amino acids),and the subcellular localization was 100% outside the cell.In addition,there were two O-glycosylation sites and abundant phosphorylation sites of NPY protein,which existed a functional domain named PAH.Spatial structure of NPY protein were mainly assembled with alpha helix and random coil.Real-time PCR results showed that the NPY gene mRNA were distributed in all tissues,there was the highest expression level of NPY gene mRNA in cerebrum,and there was the secondry highest expression level of NPY gene mRNA in pancreas and glandular stomach,which were extremely significant difference with others tussues (P<0.01),and there was the lowest expression of NPY gene mRNA in gizzard.The results would provide references for further research on the physiological regulatory functions of NPY gene on energy balance,growth and development,fat deposition,reproductive performance in poultry.

Key words: Tianzhu muscovy; NPY gene; cloning; bioinformatics analysis; tissues expression

CLC Number:

TAN Bin, YANG Shenglin, YANG Rucai, ZHOU Xuan, YANG Shihao, LUO Linli, ZHU Yongcai. Cloning and Expressing Analysis of NPY Gene of Different Tissues in Tianzhu Muscovy[J]. , 2019, 46(4): 967-976.

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Cloning and Expressing Analysis of NPY Gene of Different Tissues in Tianzhu Muscovy

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