猪NMMHC-ⅡA基因功能区的克隆及表达特征分析

畜牧兽医学报

猪NMMHC-ⅡA基因功能区的克隆及表达特征分析

蒋成兰¹，邢凤^1,2，徐云华³，王猛¹，李艳平¹，康丽¹，姜运良^1*，周恩民^1,4

（1. 山东农业大学动物科技学院，山东省动物生物技术与疫病防治重点实验室，泰安 271018； 2. 塔里木大学
动物科学学院，阿拉尔 843300； 3.莱芜市畜牧兽医技术推广中心，莱芜 271100; 4.西北农林科技
大学动物医学院，杨凌 712100）

收稿日期:2011-11-15 出版日期:2012-10-25 发布日期:2012-10-25
通讯作者: 姜运良，E-mail: yljiang723@yahoo.com.cn
作者简介:蒋成兰（1986-），女，山东费县人，硕士生，主要从事动物分子遗传学研究，E-mail: jiangchenglan@gmail.com
基金资助:
国家转基因专项（2009ZX08009-147B）；国家科技支撑计划（2008BADB2B01）；山东省良种工程（2007LZ013-03）

Cloning and Homology of the Functional Region of Porcine NMMHC-ⅡA Gene and Its Expression Pattern in Pigs

JIANG Cheng-lan¹, XING Feng^1,2，XU Yun-hua³, WANG Meng¹, LI Yan-ping¹, KANG Li¹, JIANG Yun-liang^1*，ZHOU En-Min^1,4

(1. Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and
Prevention, College of Animal Science and Technology, Shandong Agricultural University, Taian
271018, China; 2. College of Animal Science, Tarim University, Alar 843300, China; 3. Application
Center for Animal Science and Veterinary Medicine Techniques of Laiwu, Laiwu 271100, China；
4. College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China)

Received:2011-11-15 Online:2012-10-25 Published:2012-10-25

摘要/Abstract

摘要： 为了解NMMHC-ⅡA是否与不同品种猪对猪繁殖与呼吸综合征病毒(PRRSV)的抗性差异有关，本研究克隆了对PRRSV易感的三元杂交猪（杜大长）、抗PRRSV的我国优良地方品种莱芜猪和大蒲莲猪NMMHC-ⅡA基因中含PRRSV结合区（23个氨基酸）在内的羧基端954 bp的cDNA序列，并分析了其在物种间的同源性和在不同品种间的差别，通过RT-PCR 法检测了该基因在猪体内的表达特征。结果表明，NMMHC-ⅡA的cDNA序列在哺乳动物中的保守性较高，猪与人、大鼠、小鼠、牛、猴、狗等物种的核苷酸及氨基酸序列同源性均较高；莱芜猪、大蒲莲猪及杜大长杂交猪NMMHC-ⅡA 蛋白羧基端的氨基酸序列完全相同，而NMMHC-ⅡA基因的核苷酸序列有3处不同。猪NMMHC-ⅡA基因在猪肝脏、肾脏、脾脏、肺脏、淋巴结、扁桃体、大肠、小肠、胃和肌肉中均有明显的表达，而在心脏和大脑中有弱表达，在小脑及子宫中几乎检测不到其表达。结果显示，对蓝耳病呈现不同抗性的杜大长杂交猪和莱芜猪、大蒲莲猪NMMHC-ⅡA基因的功能区没有区别，该基因在除小脑和子宫外的多种组织中均表达，NMMHC-ⅡA与猪抗蓝耳病的关系需进一步研究。

Abstract: To investigate whether NMMHC-ⅡA was related to the resistance to PRRSV, the partial region of NMMHC-ⅡA gene 954 bp encoding the C-terminal region of NMMHCⅡA protein including 23 amino acid residues bound to PRRSV in hybrid pigs(Duroc×Yorkshire×Landrace), Laiwu and Dapulian pigs was cloned; then its homology with other species，the differences at nucleotide and amino acid sequences level between pig breeds and mRNA expression pattern in porcine tissues were analyzed. The results showed that NMMHC-ⅡA cDNA sequence was highly conservative among mammalian species, NMMHC-ⅡA had high homology with human, rat, mouse, cattle, monkey and dog, respectively at nucleotide and amino acid levels. Alignment of nucleotide sequence of NMMHC-ⅡA gene among Laiwu, Dapulian and Duroc×Yorkshire×Landrace crossbred commercial pigs revealed three nucleotide changes, however, the amino acid sequence was identical. NMMHC-ⅡA mRNA expression was analyzed in different tissues by RT-PCR, its expression level was high in skeletal muscle, lung, liver, kidney, spleen, bronchial lymph node, tonsil, stomach, large intestine, small intestine tissues, low in heart and cerebrum, and nearly undetectable in cerebellum and uterus. The results reveal that there is no differences in the function of NMMHC-ⅡA gene among Laiwu, Dapulian and Duroc×Yorkshire×Landrace crossbred commercial pigs, and it is expressed in many tissues except cerebellum and uterus. The role of NMMHC-ⅡA in resistance to PRRSV requires further study.

中图分类号:

S828
S813.3

蒋成兰，邢凤，徐云华，王猛，李艳平. 猪NMMHC-ⅡA基因功能区的克隆及表达特征分析[J]. 畜牧兽医学报, doi: .

JIANG Cheng-lan, XING Feng，XU Yun-hua, WANG Meng, LI Yan-ping. Cloning and Homology of the Functional Region of Porcine NMMHC-ⅡA Gene and Its Expression Pattern in Pigs[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, doi: .

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