山羊KLF17基因克隆与组织表达

Cloning and tissue expression of goat (Capra Hircus) KLF17 gene

克隆山羊Krüppel样因子17基因(KLF17)的c DNA序列,并检测其组织表达水平,为KLF17基因的功能研究奠定基础.随机选取2~3周岁的健康简州大耳羊和藏山羊羯羊各4只,屠宰采集山羊不同组织样品(心、肝、脾、肺、肾、肌肉和皮下脂肪),提取组织总RNA,利用RT-PCR法克隆山羊KLF17基因序列,进行生物信息学分析,并利用实时荧光定量PCR(q PCR)法检测KLF17基因mRNA的表达水平.结果表明,克隆得到山羊KLF17基因c DNA序列1 450 bp,包含CDS区全长858 bp,5'UTR序列12 bp,5'UTR序列580 bp,编码285个氨基酸残基.其CDS区与牛、绵羊相比在5'端少99 bp,而在531位多出361 bp,这导致其编码区在858位提前终止.山羊蛋白序列较牛、绵羊少113个氨基酸残基.组织表达结果显示,KLF17基因在藏山羊肺中具有最高的表达水平,而在简州大耳羊皮下脂肪中具有最高的表达水平.两种山羊均在背最长肌中具有最低的KLF17表达量.这些结果表明KLF17可能在山羊脂质代谢过程中具有重要调控作用.

The aim of the study was to clone the cDNA of goat KLF17 gene, and to determine its expression in various tissues, which might provide basis for research of its function in regulating lipid metabolism in goat. Each of 4 healthy ,2-3 years old cas- trated Jianzhou Big-eared goats and Tibetan goats were selected. After slaughter, the tissue samples from heart, liver, spleen, lung, kidney,longissimus dorsi muscle and subcutaneous fat were collected for total RNA extraction. The sequence of KLF17 gene was cloned by RT-PCR and analyzed using bioinformatics. The mRNA expression of KLF17 gene was determined using re- al-time fluorescent quantitative PCR（qPCR）. A length of 1 450 bp cDNA of KLF17 gene was cloned, containing 858 bp CDS, 12 bp 5＂ UTlt and 580 bp 3 ＂UTR, encoding 285 amino acids. Comparing with the CDS of cattle and sheep, goat KLF17 was shor- ter by 99 bp at 5＂UTR,and longer by 361 bp at the position of 531 bp,which resulted in pre-termination of goat CDS at 858 bp. The protein sequence of goat KLF17 was shorter by 113 amino acids compared with that of cattle and sheep. The results of tissue expression showed that the expression of KLF17 was higher in lung of Tibetan goat and subcutaneous fat of Jianzhou Big- eared goats. The longissimus dorsi muscle expressed the lowest level of KLF17 mRNA in both Tibetan goats and Jianzhou Big- eared goats. These results imply that KLF17 gene may play an important role in regulating lipid metabolism in goat.