山羊clock和cry1基因的克隆及其在大脑和垂体中表达差异的研究

畜牧兽医学报

山羊clock和cry1基因的克隆及其在大脑和垂体中表达差异的研究

占思远¹，罗万伟¹，程波¹，蒋婧¹，李利¹，王林杰¹，张红平^1*，王永²，龚华斌³，邓中宝³

（1. 四川农业大学动物遗传育种研究所，雅安625014； 2. 西南民族大学，成都 610225；3. 四川省简阳大哥大牧业有限公司，简阳 641421)

收稿日期:2012-02-28 出版日期:2012-11-26 发布日期:2012-11-26
通讯作者: 张红平，教授， E-mail：zhp@sicau.edu.cn
作者简介:占思远（1987-）,男,湖北黄冈人,硕士生,主要从事动物遗传育种与繁殖研究,E-mail:zzssyy322@163.com；罗万伟（1990-）,男,四川宜宾人, E-mail: luowanwei2010@163.com。二者共同为第一作者
基金资助:
四川省教育厅重点项目（08ZA077）；四川省科技支撑计划（2011NZ0003；2012NZ0008）

Molecular Cloning and Differential Expression of clock and cry1 Genes in Goat Brain and Pituitary

ZHAN Si-yuan¹，LUO Wan-wei¹，CHENG Bo¹，JIANG Jing¹，LI Li¹，WANG Lin-jie¹，ZHANG Hong-ping^1*，WANG Yong²，GONG Hua-bin³，DENG Zhong-bao³

(1. Institute of Animal Genetics and Breeding，Sichuan Agricultural University，Ya’an 625014, China； 2. Southwest University for Nationalities, Chengdu 610225, China; 3.Jianyang Da-ge-da Animal Husbandry Limited Company of Sichuan Province, Jianyang 641421, China)

Received:2012-02-28 Online:2012-11-26 Published:2012-11-26

摘要/Abstract

摘要： 本研究旨在探讨生物钟基因clock和cry1在山羊出生后不同时期的大脑和垂体中的表达差异。在出生后30、60、90和120 d共屠宰24只南江黄羊羔羊（公母各12只），取其大脑皮质层和垂体组织样品用于抽提RNA，进行分子克隆和实时荧光定量PCR分析。结果，分离克隆得到了山羊clock基因外显子1到外显子8以及cry1基因外显子6到外显子15 的CDS区序列，其长度分别为1 479 bp和993 bp，其中clock基因部分CDS区序列与绵羊、牛、人和小鼠的对应序列相似性分别为99%、90%、94%和92%，cry1基因部分CDS区序列与绵羊、牛、人和小鼠的对应序列相似性分别为99%、98%、94%和87%。组织表达分析结果表明，性别对clock和cry1基因的表达影响不显著（P>0.05），2个基因在垂体中的mRNA表达量极显著高于大脑中的表达量（P<0.01）；在不同时期的垂体组织中，clock和cry1基因表达量均呈现先上升后下降的趋势；在不同时期的大脑组织中，clock基因的表达量趋于波动平衡，呈现上升趋势，cry1基因的相对表达量呈细微的上升趋势。结果提示：clock和cry1基因的CDS区在物种间保守性较高。clock和cry1基因在山羊大脑和垂体组织中起到重要的作用，且其mRNA发育性表达模式具有组织特异性。

Abstract: The objective of the present study was to investigate the differential expression of clock and cry1 genes in the brain and pituitary of postnatal goats. The samples of cerebral cortex and pituitary were collected from a total of 24 (12 males and 12 females) Nanjiang Yellow goats at 30，60，90 and 120 days of age after birth. The mRNA levels of clock and cry1 genes were detected by real-time fluorescence quantitative PCR. The partial CDS of clock and cry1 genes were obtained by RT-PCR, containing exon 1 to exon 8 in clock gene and exon 6 to exon 15 in cry1 gene, and the lengths were 1 479 and 993 bp，respectively. The nucleotide sequence of CDS shared 99%, 90%, 94% and 92% consistency with the clock gene of sheep, cattle, human and mouse, respectively. Whereas the nucleotide sequence of CDS shared 99%, 98%, 94% and 87% consistency with the cry1 gene of sheep, cattle, human and mouse. The mRNA levels of clock and cry1 genes in pituitary were significantly higher than that in brain（P<0.01）, however there was no significant difference（P>0.05） between sexes. In pituitary, the mRNA levels of clock and cry1 genes were upregulated from 30 to 60 days, and then decreased. In addition, the expression of clock gene showed no significant difference at four development stages of brain, but the cry1 gene was upregulated slightly from 30 to 120 days. These results indicate that clock and cry1 genes are highly conservative, and play important roles in the brain and pituitary in goats. The mRNAs express developmentally in a tissue-specific manner.

中图分类号:

S827.2

占思远，罗万伟，程波，蒋婧，李利，王林杰，张红平，王永，龚华斌，邓中宝. 山羊clock和cry1基因的克隆及其在大脑和垂体中表达差异的研究[J]. 畜牧兽医学报, doi: .

ZHAN Si-yuan，LUO Wan-wei，CHENG Bo，JIANG Jing，LI Li，WANG Lin-jie，ZHANG Hong-ping，WANG Yong，GONG Hua-bin，DENG Zhong-bao. Molecular Cloning and Differential Expression of clock and cry1 Genes in Goat Brain and Pituitary[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, doi: .

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