一个棉花类受体蛋白激酶基因的克隆与表达分析

摘要

The cDNA encoding the full length of a receptor-like protein kinase of cotton was cloned through degenerate primers amplification, genome walking and RT-PCR.Sequence analysis showed that introns were absent in the gene.It contains a 2964 bp open reading frame, encodes a deduced protein of 988 amino acid residues, and shows 60％ homology to HAESA-like2,which regulates floral organ abscission in Arabidopsis.This gene was tentatively designated as GRLK5.GRLK5 expresses higher in seed, stem bark, root, and fibre than in other organs.The ABA treatment induce the expression of GRLK5.The over expression vector was constructed by inserting GRLK5 into the pCAMBIA2301 , tobacco plants were transformed by co-cultivating leaves method via Agrobacterium mediation.The object gene was verified to have been integrated into the genome of tobacco by PCR.%利用简并引物扩增、染色体步移以及RT-PCR的方法,分离了棉花的一个类受体蛋白激酶基因的全长序列.序列分析表明,该基因没有内含子,开放读码框的长度为2964 bp,编码的多肽序列含有988个氨基酸并与拟南芥控制花器官脱落的基因HAESA-like2具有60%左右的相似性,将该基因命名为GRLK5.半定量RT-PCR分析发现GRLK5在种子、茎皮、根和纤维中的表达量较高.并且受ABA的诱导表达.将GRLK5整合到植物表达载体pCAMBIA2301中,构建植物过量表达载体,通过农杆菌介导,采用叶盘法转化烟草,PCR检测证明目的基因已经整合到烟草基因组中.