两株蛋白核小球藻 rbcS cDNA 全序列的克隆和分析

以2株蛋白核小球藻(Chlorella pyrenoidosa)F-9和820为实验材料,根据已知绿藻rbcS基因设计简并引物,分别克隆到2条245bp的cDNA片段,以此片段为基础使用cDNA末端快速扩增(RACE)技术,获得了2条长度分别为861bp和907bp的rbcS基因。序列分析表明,蛋白核小球藻F-9和820的rbcS分别编码181和184个氨基酸,编码区具有与高等植物rbcS保守序列YYDGRYWTMWKLPMFG相类似的结构,起始密码子附近有Kozak保守序列(A/GXXATGG),3′非翻译区有加尾信号TGTAA。同源性分析结果表明,F-9与蛋白核小球藻(GenBank登录号BAE48226)的相似性最高(91%),而F-9与820、F-9和普通小球藻(C.vulgaris)的相似性分别为64%和50%。这2条rbcS基因与其他绿藻和高等植物也表现了广泛的同源性,但相似性不高。该研究旨在为rbcS基因的功能和表达研究奠定基础。

Cloning and analysis of rbcS full-length cDNA sequences from two strains of Chlorella pyrenoidosa

Ribulose-1,5-bisphosphate carboxylase/oxygenase(Rubisco)is one of the key enzymes catalyzing the CO 2 fixation in the Calvin cycle.RbcS,the small subunit of Rubisco,plays a role in regulating the Rubisco activity.In this study,two rbcS genes of unicellular green algae Chlorella pyrenoidosa strains F-9 and 820 were cloned and analyzed, respectively.Two cDNA sequences of 245 bp were obtained according to a pair of degenerate primers,and then two full-length rbcS cDNA sequences were cloned by rapid amplificati...