蒙古绵羊β-防御素-1(SBD-1) cDNA的扩增与组织表达

The Amplifying of the cDNA and Tissue Expression of β-defensin-1 Gene of Mongolian Sheep

为了获得蒙古绵羊β-防御素-1(SBD-1)的cDNA序列,从蒙古绵羊瘤胃的上皮组织中提取总RNA,采用反转录PCR(RT-PCR)技术扩增蒙古绵羊SBD-1的部分cDNA,结果扩增出cDNA 300个碱基.该cDNA包含由192个碱基组成的开放读码框(ORF),该ORF编码64个氨基酸的前原防御素.为了检测SBD-1mRNA在蒙古绵羊体内可能的表达器官,通过RT-PCR方法研究了SBD-1基因在蒙古绵羊各组织中的表达分布,结果显示,该基因在整个消化道(除直肠外)、气管、子宫、脾脏等管状器官内的黏膜层和脾脏内均有表达,而在所检测的实质性器官内,如心脏、肝脏、肺脏、肾脏、淋巴结、膀胱、卵巢等中没有表达.

To amplify and sequence the cDNA encoding sheepβ-defensin-1（SBD-1） by the reverse transcription PCR（RT-PCR）,the total RNA is extracted from the rumen epithelial tissue of mongolian sheep.The sequencing result showed that the amplified cDNA were 300 base pairs containing an open reading frame（ORF） of 192 base pairs encoding 64 amino acid prepro-β-defensin with consensus sequence of six invariantly spaced cysteine residues.RT-PCR for RNA extracted in different tissues of the Mongolian sheep indicated that SBD-1 expressed in the trachea,uterus,spleen and mucosa of the most of digestive tract（tongue,esophagus,rumen,abomasum,reticulum,omasum,duodenum,jejunum,ileum,colon） except rectum,but SBD-1 mRNA were not detected in the heart,liver,lung,kidney,lympho-node,bladder and ovarium.