中文摘要：

本研究旨在对山羊溶酶体α-甘露糖苷酶（α-AMA）基因进行组织表达谱和生物信息学分析。参考牛α-AMA基因序列设计引物,采用PCR技术克隆山羊α-AMA基因序列,并利用荧光定量RT-PCR进行组织表达谱分析以及进行生物信息学预测。首次获得了山羊α-AMA基因,含有完整CDS编码区3 000bp,编码999个氨基酸,其中前50个氨基酸为信号肽序列。其编码区的核苷酸序列和预测氨基酸序列与牛的α-AMA相似性最高,分别为95.93%和94.79%。组织表达谱分析表明α-AMA在山羊各组织均不同程度的表达,其中在肺脏、肝脏、小脑表达量较高。生物信息学预测发现,α-AMA蛋白属于糖苷水解酶38家族成员,有2个保守的结构域,存在9个N-糖基化位点。SWISS-MODEL同源建模山羊α-AMA具有良好的可信度。本研究为探讨酶的作用机理及疯草解毒剂的研制提供了理论依据。

英文摘要：

Goat lysosomal α-AMA gene was amplified using RT-PCR and the tissue specific expression profile,bioinformatics characteristic of α-AMA were studied.Primers were designed based on the sequence of bovine lysosomal α-AMA gene and were used in amplifying the goat α-AMA,the tissue specific expression profile was analyzed by qRT-PCR,and the bioinformatics analysis of α-AMA was conducted.The results showed that CDS sequence of goat α-AMA was 3 000 bp,encoding a deduced protein containing 999 amino acid residues in which the first 50 residues were signal peptide,and this nucleotide sequence of CDS and the deduced amino acids shared 95.93% and 94.79% homology with the α-AMA mRNA of cattle.The qRT-PCR revealed that the α-AMA gene was expressed in various tissues at different levels.The expression level of this gene was higher in the lung,liver,and cerebellum.It was predicted that α-AMA was belonged to Glycosyl hydrolases family 38,and composed of two conserved domains.There were thirty-five phosphorylation sites,one phosphorylation site of specific protein kinase and nine N-glycosylation sites in α-AMA protein.By SWISS-MODEL homologous modeling,the predicted structure of goat was similar to the template and predictive.These results provide the theoretic basis for the mechanism research of α-AMA and development of toxin antidote toward locoweed.