蒙古冰草Actin基因片段的克隆及序列分析

摘要

In this study,Actin gene homology fragment from Mongo1ian wheatgrass(Agropyron mongolicum Keng)was isolated using homology-based method, so that it can be used as internal standard when study other genes' expression and regulation in Mongo1ian wheatgrass.Two pairs of primers A4 and A5 was designed according to the amino acid conserved regions of the cloned actin gene of Gramineae plant wheat, and two partials gene sequences were obtained by reverse transcription polymerase chain reaction(RT-PCR),the product of which was named as MwACT.The sequences was 962 bp after merging two partials gene repeat sequences,which encoded 237 amino acids. Homology comparison with Actin gene sequences in other plants was over 79%,which was 94% identical to wheat and barley.Over 90%amino acid sequence homology was showed with other plants.%旨在利用同源序列法分离蒙古冰草(Agropyron mongolicum Keng)Actin基因同源片段,为研究其他基因在蒙古冰草中的表达和调控提供内标参照.根据禾本科植物小麦Actin基因(AB181991)的保守序列设计2对引物A4和A5,采用RT-PCR扩增蒙古冰草的Actin基因片段,分别得到656 bp和848 bp的片段,使用DNAman和DNAuser等分子生物学软件进行序列分析,将2个片段的重复序列合并后获得一段长度为962 bp的基因片段,编码237个氨基酸,将克隆的Actin基因片段命名为MwACT.该序列与其它植物Actin基因核苷酸序列的同源性均在80%以上,其中与小麦、大麦的同源性达到94%;与氨基酸序列的同源性均在90%以上.