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作 者:孙雨婷[1] 罗军[1] 朱江江[1] 赵旺生[1] 李君[1] 钟瑜[1] 郝娟[1]
机构地区:[1]西北农林科技大学动物科技学院陕西省农业分子生物学重点实验室,杨凌712100
出 处:《畜牧兽医学报》2012年第2期319-323,共5页ACTA VETERINARIA ET ZOOTECHNICA SINICA
基 金:转基因生物新品种培育项目(2009ZX08009-162B);陕西省重大科技创新项目(2009ZKC07-01-1);公益性行业(农业)科研专项(201103038)
摘 要:旨在克隆奶山羊GPR41(G protein-coupled receptor 41)基因并分析其组织表达谱,为进一步探讨其功能奠定基础。根据GenBank上已登录的牛、人和鼠的GPR41基因序列设计1对特异性引物,采用RT-PCR方法克隆奶山羊GPR41基因,利用实时荧光定量PCR方法分析奶山羊GPR41mRNA表达的组织特异性。测序结果表明奶山羊GPR41基因的CDS区为978bp,共编码325个氨基酸。奶山羊GPR41基因序列同源性分析表明:其与牛、人和鼠的核苷酸序列同源性分别为96%、78%和74%,与牛、人和鼠的氨基酸序列同源性分别为97%、76%和74%。实时荧光定量PCR分析结果表明:奶山羊GPR41基因在小肠组织中表达量最高,其次是乳腺组织,在心脏和肾脏中表达量极低。试验结果表明GPR41可能在小肠和乳腺组织中发挥着重要的生理作用。The aim of this study was to lay a foundation for understanding the physiological func- tions of dairy goat GPR41 gene by cloning GPR41 gene and analyzing its tissue expression level. Based on the published nucleotide sequence of GPR41 gene of bovine, human and mouse in Gen- Bank, a pair of special primer was designed. The dairy goat GPR41 gene was amplified by RT- PCR, the tissue specificity of dairy goat GPR41 gene were analyzed by real-time fluorescence quantitative PCR. The results showed that the coding region length was 978 bp, coding 325 ami- no acids. The sequence homology analysis of dairy goat GPR41 gene indicated that the homology of GPR41 of dairy goat was 96%, 78% and 74% compared with that of bovine, human and mouse, while the amino acid sequence homology was 97%,76% and 74% , respectively. The ex- pression analysis of GPR41 gene revealed that dairy goat GPR41 gene mRNA is expressed abun- dantly in intestine tissue and, to a lesser degree, in mammary gland, but lowly in heart and kid- ney. These results suggest that GPR41 may play an important role in intestine and mammary gland tissue.
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