蒙古红鲌肥胖基因cDNA的克隆与组织表达特异性研究  

THE MOLECULAR CLONING OF OBESE cDNA OF ERYTHROCULTER MONGOLICUS AND ITS ORGANIZATION EXPRESSION SPECIFICITY

作　　者：潘庭双[1] 龙良启[1] 吴小平[1] 罗晓松[2] 邹世平[2] 

机构地区：[1]华中农业大学水产学院,武汉430070 [2]中国水产科学研究院长江水产研究所,荆州434000

出　　处：《水生生物学报》2007年第1期94-98,共5页Acta Hydrobiologica Sinica

基　　金：湖北省自然科学基金(NO:2000JJ04)资助

摘　　要：本文根据已知的人和鼠ob基因序列设计引物,采用RT-PCR(reverse transcriptase-polymerase chain reaction)技术,从蒙古红脂肪组织RNA中扩增到ob基因片段,获得ob基因编码区438bp的序列。与人和其他动物比较,蒙古红与人、猪和鼠ob基因核苷酸编码序列的同源性分别为82.9%、84.0%和99.1%,蒙古红ob基因编码的蛋白leptin氨基酸序列与人、猪和鼠的同源性分别为80.8%、78.8%和95.9%;运用RT-PCR技术分析ob基因表达的组织特异性,结果表明:ob基因在脂肪和肝脏组织中表达量最大,在心脏、脾脏、肌肉、脑、卵巢中表达量减少,在肾脏中微量表达,而在精巢和肠道组织中不表达。In this study, Erythroculter mongolicus were taken from the Nanhu Lake, the length and weight of male and female were about 16 centimeters, 112 grams and 18 centimeters, 120 grams, respectively. Tissues such as adipose, liver, heart, spleen, muscle, brain, ovary, kidney and spermary had been obtained from the fish and reserved. After washed with distilled water, total RNAs of each tissue samples were extracted by one step method and the extracted RNA was identified by Eppendorf Bio-photometer. All the total RNA that ratios of OD280/OD260 were diluted to 0.5μg/μL. The primers designed according to the consensus region of human and mouse ob genes were employed to perform RT-PCR （reverse transcriptase-polymerase chain reaction）. The product of RT-PCR from total RNA of E. mongolicus fat tissue and pBluescript SK（ ＋ ） vector were both digested by EcoR Ⅰ and Hind Ⅲ , then linked and transformed into E. coli competent cell. The positive recombinant plasmid was sequenced and got a 438bp coding region DNA sequence. Compared to the nucleotide sequences of ob genes in human, pig and mouse, the ob genes of E. mongolicus had a homology as 82.9%, 84.0% and 99.1%, respectively. The nucleotide sequences of ob genes were very conservative among the four animal species. In comparison in the leptin amino acid sequence with human, pig and mouse, the homologies were 80.8 %, 78.8 % and 95.9 % respectively. The result of ob gene tissue express specificity by RT-PCR method indicated that ob gene was expressed mainly in adipose and liver, less in heart, spleen, muscle, brain, ovary, slightly in kidney and none in spermary and intestines in E. mongolicus.

关 键 词：蒙古红鲌 OB基因 LEPTIN 组织 

分 类 号：Q953[生物学—动物学]