德州驴朊病毒基因克隆与序列分析

根据已报道正常哺乳动物朊蛋白(PrP)基因序列设计引物,采用PCR法扩增了德州驴的PrP基因,将其克隆到T-Vector。序列测定及分析表明所克隆的德州驴PrP基因片段为767 bp,该基因内无内含子,包含了驴PRNP完整编码区序列,编码256个氨基酸的前体蛋白,推测其分子量约34 ku。通过对德州驴朊病毒核苷酸和氨基酸序列与其他7种动物的比较,我们发现有3个区域变异较大,分别与种属屏障和潜伏期有关。经推测驴的PrP基因型是ARQ型,对羊痒病中度敏感,但是至今没有马属动物感染羊痒病的报道。这为TSE种间传播的突破提供了基础数据。

Cloning and sequence analysis of prion gene of the Shandong dezhou donkey

Based on the reported prion gene sequence of mammal, we designed the primers.Using the PCR technique,we obtained the prion gene of Dezhou donkey and cloned it into T-vector.The sequencing analysis showed that the gene was 767bp and had no intron.It included the intact ORF and coded the former prion protein that had the 256 amino acids.We inferred that the molecular weight of donkey prion protein was 34ku.Comparing the prion gene nucleic acid sequence and AA composion,we analyzed the epitope of species barrier of prion disease.It can be confirmed that the genotype of the donkey was ARQ and medium sensitive to scrapie.However,there is no report about equus being infected with scrapie.All this will provide the basic data of species transmission for TSE.