辽宁绒山羊UMP/CMP激酶基因的表达及分析

摘要

In order to study the genetic characteristics of Liaoning newbreed cashmere goat,on the basis of construction of a skin cDNA library and isolation of a full-length cDNA clone termed UMP/CMPK,the bioinformatics analysis and semi-quantitative RT-PCR were used to detect the expression of the UMP/CMPK gene in all internal organs. A bioinformatics analysis showed that the UMP/CMPK gene encoded a putative 228 amino acid protein that exhibited a high content of leucine,glycine and lysine and had a lower content of histidine and tryptophan. The secondary structure prediction revealed a high proportion of random coils (51.75%). UMP/CMPK contained a conservative UMP/CMP kinase protein structure domain. UMP/CMPK of the goat and cattle had the highest homology. UMP/CMPK also had transmembrane domain and signal peptide. 8 Ser,2 Thr and 2 Tyr may be phosphorylation sites in CMP/UMPK protein.RT-PCR analysis showed that UMP/ CMPK were only present in goat skin. This research provides an experimental basis for the further study of the genetic characteristics of the goat.%在前期构建辽宁绒山羊皮肤cDNA文库的基础上,测通得到了新基因UMP/CMP激酶的全长序列,并对UMP/CMP激酶基因进行生物信息学分析.然后,通过RT-PCR检测该基因在心、肝、脾、肺及肾中的表达.生物信息学分析显示,UMP/CMP激酶基因全长1 081 bp,能够编码含有228个氨基酸的蛋白质,亮氨酸、甘氨酸、赖氨酸含量较高.UMP/CMP激酶含有较高比例的无规卷曲结构(51.75％),并发现其含有UMP/CMP激酶蛋白结构域.蛋白三级结构、进化树分析显示,辽宁缄山羊和牛的UMP/CMP激酶基因的同源性最高.UMP/CMP激酶基因存在跨膜结构域,并且存在信号肽剪切位点.该基因中有8个Ser,2个Thr,2个Tyr可能成为蛋白激酶磷酸化位点.RT-PCR检测显示,UMP/CMP激酶基因特异地表达于皮肤.这些研究结果为深入研究辽宁绒山羊的种质遗传特性奠定了基础.