杜氏盐藻RuBisCO小亚基基因的克隆和分析  

Analysis and Cloning of the Small Subunit Gene of RuBisCO of Dunaliella salina

作　　者：柴玉荣[1] 刘国红[2] 刘红涛[1] 李杰 薛乐勋[1] 

机构地区：[1]郑州大学细胞生物学研究室,中国河南郑州450052 [2]郑州大学基础医学院组胚教研室,中国河南郑州450052

出　　处：《生命科学研究》2008年第1期29-34,共6页Life Science Research

基　　金：国家自然科学基金资助项目(30540067)

摘　　要：根据莱茵衣藻(Chlamydomonas reinhardtii)、团藻(Volvox carteri)、伞藻(Acetabularia cliftonii)等生物的1,5-二磷酸核酮糖羧化酶/加氧酶(RuBisCO)的小亚基rbcS基因氨基酸的高度保守序列,设计一对简并引物,进行RT-PCR.209bp的PCR产物经测序分析及进行氨基酸序列同源性比对,表明克隆的序列为盐藻rbcS基因的cDNA片段.根据该序列信息,采用RACE(rapid amplification of cDNA ends)的方法扩增其5′上游未知区和3′下游未知区.5′RACE得到的cDNA长度为约300bp,3′RACE得到的长度约380bp.三段序列拼接后cDNA全长为878bp,其中开放读码框包括190个氨基酸.此cDNA序列,推导成氨基酸序列与已知物种的rbcS基因相比对,同源性分别为V.carteri78%,C.reinhardtii75%,A.cliftonii67%,据此可推断所克隆的序列为盐藻RuBisCO的小亚基cDNA序列,GenBank收录号为AY739272.Ribulose-1,5-bisphosphate carboxylase/oxygenase （Rubiscr） is a key enzyme in photosynthesis.A full-length cDNA encoding the rbcS of D.salina was cloned by the reverse transcription polymerase chain reaction （RT-PCR） and rapid amplification of cDNA ends （RACE） were reported. One pair of degenerate primer was designed according to conserved motifs of known rbcS from .Chlamydomonas reinhardtii, Volvox carteri,Acetabularia clifionii etc, and was used to amplify the rbcS cDNA fragment from the total RNAs of D.salina. The resulting cDNA was 209 base pairs in length and shared high identify with other known rbcS. According to the rbcS cDNA sequence,5＇ RACE and 3＇ RACE were performed to obtain the 5＇ upstreaming and 3＇ downstreaming sequences of the rbcS gene.The total length of the three fragments was 878 bp which contained an open reading frame encoding 190 amino acids. Homologous analysis indicated that the deduced amino acid sequences of rbcS had a high degree of identity with previously reported members of other rbcS gene. The nucleotide sequence data reported here had appeared in the GenBank database under accession number AY739272.

关 键 词：盐藻 1 5-二磷酸核酮糖羧化酶 加氧酶 RBCS RACE 

分 类 号：Q754[生物学—分子生物学]