结缕草肌动蛋白基因全长cDNA的克隆及序列分析

摘要/Abstract

摘要： 根据植物肌动蛋白(Actin)基因编码区的保守序列设计引物,提取结缕草叶片的总RNA,进行RT-PCR。并采用RACE技术扩增出1560bp的Actin基因全长cDNA序列。序列分析表明,该基因的开放阅读框(ORF)为1134bp,编码377个氨基酸,5′非编码区117bp,3′非编码区309bp。所得序列与GenBank中收录的其他植物肌动蛋白核苷酸序列的一致性均在85%以上,氨基酸序列的一致性高达97%以上。将其命名为ZjACT,GenBank登录号为GU290545。根据高等植物肌动蛋白相似性构建的系统进化树显示,结缕草肌动蛋白与大麦和圆锥小麦肌动蛋白之间的亲缘关系最为密切,在进化中分化时间最为接近。进一步分离克隆了结缕草Actin基因的基因组DNA序列(登录号GU290546),它由4个外显子和3个内含子组成。本研究有助于揭示植物Actin基因家族的进化历史,为研究植物Actin基因家族功能和进化上的多样性奠定理论基础,同时也为开展草坪草和牧草Actin基因的功能分析和利用研究提供参考。

Abstract: One 1 560 bp full-length cDNA of Actin was cloned from leaves of zoysia grass using RT-PCR and RACE methods. The sequence analysis revealed that it contained an ORF of 1 134 bp encoding a protein composed of 377 amino acids, a 5′-UTR of 157 bp and a 3′-UTR of 495 bp. Homologous alignment showed that it shared over 85% of nucleotide identities and 97% of amino acid identities with actins from other plants in GenBank. The full-length cDNA was designated as ZjACT with its accession number GU290545. The phylogenetic tree constructed on the basis of amino acid sequences suggested that the relationship of actins from zoysia grass was most intimate with that from barley and poulard wheat, and they might share the same differential time in evolution. A comparison of the cDNA and its genomic counterpart (GU290546) demonstrated that the gene consisted of 4 exons and 3 introns. These results provided clues for evolutionary studies of the actin gene family, and on its functional and evolutionary diversity, as well as served as references for functional analysis and utilization of Actin gene from turf and forage grasses.

中图分类号:

王舟,宗俊勤,宣继萍,郭爱桂,刘建秀. 结缕草肌动蛋白基因全长cDNA的克隆及序列分析[J]. 草业学报, 2010, 19(6): 154-163.

WANG Zhou, ZONG Jun-qin, XUAN Ji-ping, GUO Ai-gui, LIU Jian-xiu. Cloning and sequence analysis of full-length cDNA of an actin gene from zoysia grass (Zoysia japonica)[J]. Acta Prataculturae Sinica, 2010, 19(6): 154-163.

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