猪CuZnSOD基因片段的克隆及不同生长阶段猪肝脏CuZnSOD基因表达的差异  

Cloning of CuZnSOD gene of pig and the difference of CuZnSOD gene in liver tissue at different growth stages of pig

作　　者：徐春兰[1] 汪以真[1] 

机构地区：[1]浙江大学饲料研究所教育部动物分子营养重点实验室,浙江杭州310029

出　　处：《中国兽医学报》2007年第3期394-398,共5页Chinese Journal of Veterinary Science

基　　金：国家"973"重点基础研究发展计划资助项目(2004CB117506)

摘　　要：从体质量1、20、40、60、90kg左右的杜长大猪的肝脏组织中提取基因组RNA，RT—PCR扩增CuZnSOD基因，获得了1务255bp的片段，以pGEM—TEasyVector为载体，将该片段克隆到大肠杆菌（E．coli）中，筛选阳性克隆测定其序列，分析表明该片段与报道的猪CuZnSODcDNA部分序列同源性达到100％，编码85个氨基酸组成的多肽，是成熟肽的一部分。以CuZnSOD基因片段的克隆为基础，构建了优化的半定量RT—PCR法，以18SrRNA为内标，研究不同生长阶段猪肝脏组织中CuZnSOD基因mRNA表达的差异。结果显示，1kg左右猪肝脏组织中CuZn—SODmRNA表达水平较高，体质量1～20kg猪的肝脏组织中CuZnSOD基因表达量呈下降趋势，体质量40kg的猪又显著上升，到生长后期即体质量90kg左右，CuZnSOD基因表达量又急剧下降。Total RNA was extracted from liver tissues of 1,20,40,60 and 90 kg pigs （Duroc X Landrace X York- shire） and copper zinc superoxide dismutase（CuZnSOD） gene mRNA was amplified using RT-PCR. A cDNA frag- ment about 255 bp in length was obtained and then cloned into pGEM-T vector. CuZnSOD gene was isolated and sequenced from the positive clones screened. Squence analysis suggested that this fragment was partial sequence of CuZnSOD cDNA and coded 85 amino acid residues. The gene homology of the fragment obtained in this study withthat of reported CuZnSOD cDNA sequence of porcine was 100%. Based on the CuZnSOD gene clone ,an optimal se-mi-quantitative RT-PCR method was successfully constructed. Using 18S rRNA as inner control,the difference of CuZnSOD gene expression was researched in different growth stages of pigs,it decreased from 1 kg to 20 kg,and then increased at the stage of from 20 kg to 40 kg and thereafter,it declined distinctly.

关 键 词：CUZNSOD 肝脏 基因克隆 RT—PCR 基因表达 猪 

分 类 号：Q78[生物学—分子生物学] S823[农业科学—畜牧学]