摘要
目的:克隆悬钩子属植物肌动蛋白基因(actin),为研究该物种中其他基因的表达和调控提供内标基因。方法:利用一对特异性引物从黑莓、悬钩子杂种和树莓品种中克隆actin cDNA片段,对其进行序列分析和半定量RT-PCR表达分析。结果:从3个品种中均获得一条783 bp actin cDNA片段,编码260个氨基酸,3条actin片段与其他植物核苷酸序列的同源性都在82%以上,与其他植物氨基酸序列同源性也均在95%以上;树莓和悬钩子杂种品种的actin核苷酸和氨基酸序列同源性均达99%,系统进化分析也发现两者亲缘关系较近些;表达分析发现actin基因在各品种不同组织中均有一定的表达量。结论:首次克隆了悬钩子属植物肌动蛋白基因actin,将来自黑莓和树莓品种的序列登录在GenBank,登录号分别为HQ439556和HQ439557。
Objective： To clone the actin gene from three Rubus spp.cultivars including blackberry（Navaho）,raspberry（Kivigold） and rubus hybrid berry（Youngberry） cultivars and so provide housekeeping gene for expression and regulation analysis of other functional genes in this species.Method： By using a pair of gene-specific primers of actin genes from other plants,three actin cDNA fragments were isolated from Navaho,Youngberry and Kivigold respectively,and sequence analysis and semi-quantitative RT-PCR analysis in different tissues were also performed.Result： Three cDNA fragments of 783 bp encoding 260 amino acid residues were obtained.Homology comparison with actin gene sequences in other plants showed that they shared over 82% nucleotide sequence homology and 95% amino acid sequence homology with other plants.Furthermore,actin amino acid sequences from Kivigold and Youngberry showed extreme similarity of 99% and phylogenic analysis also showed that the actin protein from Kivigold and Youngberry had close genetic relationship.Semi-quantitative RT-PCR analysis showed that actin genes in three Rubus spp.cultivars all exhibited certain level.Conclusion： It was the first report on actin genes from the genus Rubus,and the two from Navaho and Kivigold were registered into GenBank（accession numbers： HQ439556 and HQ439557）.
出处
《生物技术》
CAS
CSCD
北大核心
2011年第2期10-15,共6页
Biotechnology
基金
国家自然科学基金项目(31000753)
江苏省农业科技自主创新基金项目(CX[10]109)
江苏省科技基础设施建设计划项目(BM2009041
BM2010458)资助
