朗德鹅3-羟基-3-甲基戊二酰辅酶A合酶基因(HMGCS1)cDNA克隆及其mRNA在填饲鹅肝组织中表达的研究  

Molecular Cloning and Expression of Goose(Anser anser) 3-hydroxy-3-methylglutaryl Coenzyme A Synthase Gene(HMGCS1) in Overfeeding Landes Goose Liver

作　　者：许静[1,2] 杨倩[1] 李进军[2] 田勇[2] 陈芳[1,2] 卢立志[1,2] 

机构地区：[1]南京农业大学,南京210095 [2]浙江省农业科学院畜牧兽医研究所,杭州310021

出　　处：《农业生物技术学报》2014年第1期79-86,共8页Journal of Agricultural Biotechnology

基　　金：国家水禽产业技术体系(No.CARS-43-2);浙江省"十二五"育种专项(No.2012C12906-18)

摘　　要：本实验旨在获得鹅(Anser anser)3-羟基-3-甲基戊二酰辅酶A合酶基因(HMGCS1)的cDNA序列,观察该基因的表达与填饲鹅肝脏生成胆固醇的相关性。选用填饲28 d朗德鹅作为实验材料,运用RACE方法扩增目的基因的cDNA序列,利用荧光定量PCR技术检测并分析HMGCS1基因在各组织中mRNA的表达,比较填饲28 d后各组织HMGCS1基因表达的变化,同时选用胆固醇试剂盒测定鹅肝中胆固醇的含量。结果显示,HMGCS1基因的cDNA全长为2 947 bp,开放阅读框编码522个氨基酸(GenBank登录号:KF425515)。荧光定量PCR结果显示,该基因在肝脏中表达显著高于其他组织(P<0.05)。填饲28 d后,填饲组肝脏胆固醇含量低于对照组(P>0.05),该基因在肝脏中的表达显著降低(P<0.05)。氨基酸序列分析表明,HMGCS1基因高度保守,与鸭(Anas platyrhynchos)、鸡(Gallus gallus)、鸽子(Columba livia)等家禽同源性较高。该基因特异性地在肝组织中表达,且该基因的表达与肝脏中胆固醇合成呈正相关。本实验结果为进一步研究HMGCS1基因的功能提供了参考资料。This experiment is conducted to study the 3-hydroxy-3-methylglutaryl coenzyme A synthase gene （HMGCS1）, to obtain the cDNA sequence, and to quest the correlation between HMGCS1 gene expression and cholesterol biosynthesis in the overfeeding goose liver. We selected landes goose（Anser anser） as experiment animal, which were needed for overfeeding 28 d. The experiments were designed to clone the whole cDNA of HMGCS1 gene by RT-PCR and RACE techniques and the expression of HMGCS1 in different tissues were analyzed by fluorescence quantitative RT-PCR. We choosed a cholesterol kit to detect the cholesterol level in goose liver. Finally we got nucleotide sequence of a full-length 2 947 bp cDNA for the HMG-CoA synthase, encoding a protein of 522 amino acids（GenBank accession No.KF425515）. The results of RT-PCR showed that the gene expression in liver was extremely higher than that of other tissues（P〈0.05）. After overfeeding for 28 d, the cholesterol in the liver decreased in the experiment group, but the decrease was not significant（P〉0.05）. Nevertheless, there was a significant decrease in the level of HMGCS1 mRNA expression in experiment group（P〈0.05）. The results of amino acid sequence analysis indicated that the gene maintained a high conservatism in evolution with ducks（Anas platyrhynchos）, chickens（Callus gallus） and pigeon（Columba livia）. The gene was mainly expressed in the liver. There was a positive correlation between the cholesterol levels and the activity of the HMGCS1 in the cholesterol biosynthetic pathway. These results may provide the basic data for further research on the structure and functions of HMGCSI gene.

关 键 词：朗德鹅 3-羟基-3-甲基戊二酰辅酶A合酶基因(HMGCS1) 胆固醇合成 填饲 基因表达 

分 类 号：S858.32[农业科学—临床兽医学]