【摘要】 目的克隆隐孢子虫Gal/GalNAc特异性凝集素P30基因并测序,对其进行生物信息学分析。方法采用聚合酶链(PCR)技术,从微小隐孢子虫基因组中扩增P30基因,然后将其克隆入pMD18-T载体,转化后挑取阳性克隆进行酶切和测序鉴定,并对获得的P30基因进行生物信息学分析。结果PCR扩增得到特异的微小隐孢子虫P30基因,测得的核苷酸序列及其推导的氨基酸序列与GenBank上提交的序列同源性分别为98%～100%和99%～100%。结论成功克隆出序列正确的微小隐孢子虫P30基因,预测的P30蛋白具有9个潜在的抗原表位,为其相关研究奠定了基础。更多还原

【Abstract】 Objective To clone the galactose/N-acetylgalactosamine-specific lectin P30 gene of Cryptosporidium parvum into the plasmid pMD18-T vector for nucleotide sequencing and bioinformatics analysis. Methods The P30 gene was amplified from the genomic DNA of C. parvum by polymerase chain reaction (PCR) and cloned into the pMD18-T vector directly. Positive clones were screened and identified by EcoRⅠ and XhoⅠ digestion and sequenced. Then the DNA sequence was analyzed by relevant bioinformatics software. Results The P30 gene was specifically amplified, and comparison analysis of the nucleotide and deduced amino acid sequences were 98%-100% and 99%-100%, respectively. Conclusion The P30 gene was successfully cloned, and there were nine predicted potential antigenic determinants for the deduced P30. These results lay the foundation for future research.更多还原