中国美利奴羊(新疆军垦型)核因子I/B基因的克隆及表达  

Characterization of Tissue Expression and Cloning of Nuclear Factor I/B Gene in Chinese Merino(Xinjiang Junken Type)

作　　者：荣恩光[1] 于磊[1] 杨华[2] 张永胜 马春萍 李辉[1] 王宁[1] 

机构地区：[1]东北农业大学动物科学技术学院,哈尔滨150030 [2]新疆农垦科学院畜牧兽医研究所,新疆石河子832000 [3]新疆西部牧业股份有限责任公司,新疆石河子832000

出　　处：《中国农业科学》2012年第21期4475-4483,共9页Scientia Agricultura Sinica

基　　金：国家转基因生物新品种培育重大专项(2009ZX08009-160B;2011ZX08009-002)

摘　　要：【目的】研究绵羊核因子I/B(nuclear factor I/B,NFIB)基因的组织表达规律,并克隆中国美利奴羊(新疆军垦型)NFIB基因的全长编码区(coding sequence,CDS区)序列。【方法】采用半定量RT-PCR的方法分析NFIB基因的组织表达谱和皮肤表达特性,利用RT-PCR扩增绵羊NFIB基因的全长CDS区、克隆测序,并进行序列分析。【结果】①NFIB基因在绵羊多种内脏器官和皮肤组织中都有着不同程度的表达,且在皮肤组织中表达较高;NFIB基因在超细毛品系体表不同部位皮肤组织中的表达水平不存在显著性差异(P>0.05);同时在6个不同品系/品种绵羊体侧部皮肤组织中的表达水平也不存在显著性差异(P>0.05);超细毛品系绵羊皮肤组织NFIB基因的表达水平在不同季节存在显著性差异(P<0.05);②绵羊NFIB基因编码区至少存在3种剪接形式,其开放阅读框(open reading frame,ORF)的长度依次为1 263、1 128和1 038 bp,分别编码420、375和345个氨基酸。【结论】中国美利奴羊(新疆军垦型)NFIB基因在皮肤组织中的表达量较高,其在超细毛品系羊体侧部皮肤组织中的表达量具有显著的季节性差异;绵羊NFIB基因至少可以编码3种不同的蛋白剪接体(proteinisoform)。【Objective】The objective of this study was to explore the expression level of nuclear factor I/B gene in various sheep tissues, and clone the full-length coding sequence （CDS） of sheep NFIB gene in Chinese Merino （Xinjiang Junken type）.【Method】Tissue expression profile of NFIB gene was detected by semi-quantitative reverse transcription PCR （SqRT-PCR）. The full-length CDS region of NFIB gene was amplified by RT-PCR, cloned into T vector, and sequenced. The sequence was analyzed by using bioinformatics software.【Result】SqRT-PCR analysis showed that the NFIB gene was ubiquitously expressed in sheep internal organs and skin with different degrees. Comparatively, it was highly expressed in skin. The skin expression of NFIB gene in super fine wool strain had significant difference in different seasons （P〈0.05）, but no significant difference in skins from various anatomic locations. There was no significant difference in the NFIB gene expression of body side skin among the 6 sheep strains/breeds （P〉0.05）. Sequence analysis indicated that the sheep NFIB gene generated three NFIB protein isoforms of 420, 375 and 345 aa by alternative splicing.【Conclusion】NFIB gene is highly expressed in the skin of Chinese Merino （Xinjiang Junken type）, and its expression in the skin of super fine wool strain has significant difference in different seasons. Sheep NFIB has at least three protein isoforms. This study not only has laid a foundation for clarifying the function of sheep NFIB gene, but also provided a theoretical basis for creating high quality merino sheep.

关 键 词：NFIB基因 基因表达 中国美利奴羊 基因克隆 蛋白剪接体 超细毛品系 

分 类 号：S826[农业科学—畜牧学]