蛋白核小球藻核酮糖-1,5-二磷酸羧化/加氧酶小亚基部分基因序列的克隆与分析  

Cloning and analysis of rbcS partial gene sequences from Chlorella pyrenoidosa

作　　者：孙雪[1] 马斌[1] 周成旭[1] 

机构地区：[1]宁波大学海洋生物工程重点实验室,浙江宁波315211

出　　处：《海洋科学》2008年第3期40-43,48,共5页Marine Sciences

基　　金：国家自然科学基金项目(30700610);浙江省教育厅项目(20040899);宁波市自然科学基金项目(2006A610084)

摘　　要：以单细胞绿藻蛋白核小球藻(Chlorella pyrenoidosa)(藻株编号为NMBluh015-1)为实验材料,利用几种绿藻中的核酮糖-1,5-二磷酸羧化/加氧酶(Rubisco)的小亚基(rbcS)的保守序列,设计简并引物,克隆到rbcS的一条cDNA(245 bp)和3条DNA序列(其编号为CP1、CP2和CP3,长度依次为1425 bp、810 bp、705 bp)。序列比较结果表明,该蛋白核小球藻rbcS cDNA核苷酸序列与普通小球藻(C.vulgaris)、杜氏藻(Dunaliella tertiolecta)(rbcS1和rbcS2)及蛋白核小球藻"太阳小球藻"株的相似性依次为93%、92%(91%)和90%,而其编码的氨基酸序列与杜氏藻相似性最高(80%);3条rbcS DNA序列与绿藻及高等植物rbcS部分序列表现了较高的同源性,其中2条DNA(CP2和CP3)部分区段同源性很高。实验结果表明扩增得到的cDNA和DNA序列为蛋白核小球藻rbcS基因。In this paper, the rbcS genes from the unicellular green alga Chlorella pyrenoidosa NMBluh015- 1 were cloned and analyzed. Methods： One pair of degenerate primers specific for rhcS was designed according to the conserved rbcS nucleotide sequences from other unicellular green algae. Then, one rbcS cDNA （245bp） and three DNA （named as CP1, CP2 and CP3 with length of 1425bp, 810 bp and 705bp, respectively） sequences were PCR amplified, cloned and sequenced from C. pyrenoidosa NMBluh015-1. Results： The nucleotide sequence of rbcS cDNA was similar to those of C. vulgaris, Dunaliella tertiolecta and ＂SUN CHLORELLA＂ strain of C. pyrenoidosa with identities of 93%, 92% （91%） and 90%, respective ly. However, the deduced amino acid of rbcS cDNA had the highest identity of 80% to D. tertiolecta. The three rbcS DNA sequences showed high homology with those of other green algae and higher plants, of which CP2 sequence shared more similar to CP3 than to CP1. Conclusion： The partial rbcS sequences from C. pyrenoidosa NMBluh015-1 were obtained.

关 键 词：蛋白核小球藻(Chlorella pyrenoidosa) 小亚基(rbcS) 克隆 同源性 

分 类 号：Q781[生物学—分子生物学]