版纳微型猪近交系ARRDC5基因亚细胞定位研究  

Subcellular Location of ARRDC5 Gene in Banna Mini-pig Inbred Line

作　　者：王配[1,2] 王淑燕[1,2] 霍金龙[1,2] 张霞[1,2] 高林青[3] 赵慧敏[3] 霍海龙[4] 曾日彬 李罗刚[1,2] 滕晓红 WANG Pei;WANG Shuyan;HUO Jinlong;ZHANG Xia;GAO Linqing;ZHAO Huimin;HUO Hailong;ZENG Ribin;LI Luogang;TENG Xiaohong(Key Laboratory of Banna Mini-pig Inbred Line of Yunnan Province,Yunnan Agricultural University,Kunming 650201,China;Faculty of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201,China;Changzhi Vocational and Technical College,Changzhi 046000,China;Teaching Affairs Department,Yunnan Vocational and Technical College of Agriculture,Kunming 650031,China)

机构地区：[1]云南农业大学,云南省版纳微型猪近交系重点实验室,云南昆明650201 [2]云南农业大学动物科学技术学院,云南昆明650201 [3]长治职业技术学院,山西长治046000 [4]云南农业职业技术学院教务处,云南昆明650031

出　　处：《云南农业大学学报（自然科学版）》2020年第1期33-39,共7页Journal of Yunnan Agricultural University:Natural Science

基　　金：国家自然科学基金项目(31660637,31460580,31660650,31160439)

摘　　要：【目的】抑制蛋白5(ARRDC5)的功能尚不清楚,前期发现其可能与公猪繁殖力有关,本研究旨在克隆猪ARRDC5基因以及确定猪ARRDC5蛋白的亚细胞定位。【方法】通过NCBI下载猪近缘物种ARRDC5基因序列,在线BLAST比对获得猪EST序列,电子克隆猪ARRDC5基因编码区及部分侧翼序列。构建真核融合表达载体并瞬时转染猪睾丸细胞系ST,利用倒置荧光显微镜检测蛋白表达。【结果】获得版纳微型猪近交系(BMI)ARRDC5基因的cDNA序列1045 bp,包含1014 bp的CDS区,编码337个氨基酸,与牛及人的同源性高。成功构建了表达载体pEGFP-C1-ARRDC5,ARRDC5蛋白主要定位于细胞质中,部分细胞核中也有分布。【结论】研究结果为进一步研究该蛋白功能奠定基础。[Purpose]The function of the arrestin domain-containing protein 5(ARRDC5)was unclear up to now.Our previous research indicated that ARRDC5 might be associated with the fecundity of boars.The aim of this study was to clone the porcine ARRDC5 gene and analyze its subcellular location.[Methods]The ARRDC5 sequences of allied species of pig were obtained from NCBI and compared by online BLAST to obtain the expressed sequence tag(EST).Subsequently we used silicon cloning strategy to gain the coding sequence and a few flanking sequences of ARRDC5.Then,the eukaryotic expression vector pEGFP-C1-ARRDC5 was constructed by Digestion-Ligation and transfected into the swine testicle(ST)cells with liposome,detection of protein expression by inverted fluorescence microscopy.[Result]The cDNA length of Banna mini-pig inbred line(BMI)ARRDC5 was 1045 bp that included an open reading frame of 1014 bp coding a protein of 337 amino acids.The BMI ARRDC5 protein sequence shared high homology with that of cattle and human.We successfully constructed the expression vector pEGFP-C1-ARRDC5,and the green fluorescent was mostly limited in the cytoplasm of ST cells,which showed that the gene products of ARRDC5 in eukaryocytes were mainly located in the cytoplasm.The cell nuclei are also expressed in some cells.[Conclusion]These results will provide a basis for the study of ARRDC5 function in swine.

关 键 词：抑制蛋白5 基因克隆 版纳微型猪近交系 载体构建 亚细胞定位 

分 类 号：S828.2[农业科学—畜牧学]