摘要
[目的]为了研究绵羊甘露聚糖结合凝集素(MBL)基因。[方法]以陶塞特羊全血基因组DNA为模板,参考GenBank上牛与人的MBL基因序列合成一对特异性引物,进行PCR扩增,并经回收重组到pBS-T载体,筛选阳性克隆。用DNAMAN软件将部分测序结果同已发表的牛和人MBL序列进行同源性比较,并通过Primer Premier5进行蛋白质氨基酸序列预测。[结果]经PCR扩增获得了549bp的特异性片段。部分测序结果同牛和人MBL序列的同源性分别高达97%和86%,且该DNA序列与牛MBL部分序列编码的氨基酸仅一个之差,证实该序列为绵羊MBL基因的部分序列。[结论]绵羊MBL基因序列的获得,不仅为绵羊MBL基因全序列的获得提供了依据,还有助于进一步揭示MBL的生理和病理功能,对绵羊优良品种的选育具有深远意义。
[Objective] The aim of the research was to study the mannan-binding lectin （MBL） gene in sheep. [Method] With the whole blood genomic DNA of Dorset sheep as template, the PCR amplification was made by a pair of specific primers synthesized by referring to the GenBank report of cattle and human MBL gene. The PCR products were recombined into pBS-T vector after recovery and positive clones were screened. The homology between part of sequencing results and the reported sequence of cattle and human MBL gene was compared by DNAMAN software and the amino acid sequences of protein were predicted through Primer Premier5 software. [Result] A specific fragment with 549 bp was obtained from PCR amplification. The homology between part of sequencing results and the sequences of cattle and human MBL gene were 97 % and 86 % respectively. There was a difference of only one amino acid in coded amino acids between the DNA sequence and part sequence of cattle MBL gene, proving that the sequence was part of sheep MBL sequences. [Conclusion]The sequence of MBL gene in sheep was obtained in the research first, which provided basis for obtaining the whole sequence of MBL gene in sheep and helped to father reveal its physiological and pathological function, which had a profound significance for breeding excellent sheep varieties.
出处
《安徽农业科学》
CAS
北大核心
2007年第20期6133-6135,共3页
Journal of Anhui Agricultural Sciences
