水产学报2011，Vol.35Issue(5)：668-674,7.DOI:10.3724/SP.J.1231.2011.17221
巴夫杜氏藻1,5-二磷酸核酮糖羧化酶/加氧酶小亚基基因的克隆和分析
Cloning and characterization of a gene encoding small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase from Dunaliella parva
摘要
Abstract
Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco)is a key enzyme in photosynthesis,and its small subunit gene rbcS can affect the carboxylation catalytic efficiency and CO2/O2 specificity of the enzyme.We cloned and described the rbcS gene and promoter from Dunaliella parva.Based on the highly conserved nucleotide regions of known rbcS, a pair of specific primers were synthesized to amplify 380 bp cDNA and 1 841 bp DNA sequence in Dunaliella parva.Then the 5’ genomic DNA and 3’ cDNA sequences were cloned by Genome walking and rapid amplification of cDNA ends (RACE) technology.Based on the sequences of the 5’- and 3’-termini, primers were synthesized to obtain the full-length genomic DNA and cDNA.The full-length rbcS cDNA contained 570 bp open reading frame (ORF) ( GenBank accession no.HQ315783 ) ,294 bp of 3’ noncoding region.The full-length rbcS genomic DNA was 2 031 bp.In addition,a 476 bp promoter was obtained.Similarity analysis revealed that the highest identity was found between D.parva and Dunaliella salina.The D.parva rbcS also showed wide similarity with other green algae.This study laid foundation for further research on the function analysis and overexpression of rbcS genes, genetic manipulation of Rubisco.In addition,high levels of expression have made the promoter of the rbcS gene an attractive candidate to drive expression of transgene.Therefore the D.parva rbcS promoter after optimization should facilitate the development of transformation system of halotolerant algae and efficient expression of transgene.关键词
巴夫杜氏藻/基因克隆/5'-上游序列分析/rbcS/1,5-二磷酸核酮糖羧化酶/加氧酶Key words
Dunaliella parva/ gene cloning/ promoter analysis/ rbcS/ Rubisco分类
生物科学引用本文复制引用
尚常花,朱顺妮,袁振宏,吕鹏梅,王忠铭..巴夫杜氏藻1,5-二磷酸核酮糖羧化酶/加氧酶小亚基基因的克隆和分析[J].水产学报,2011,35(5):668-674,7.基金项目
广东省中国科学院全面战略合作项目(2010A090100010) （2010A090100010）
科技部"十二五"支撑计划(2011BAD14B03) （2011BAD14B03）
中国科学院可再生能源与天然气水合物重点实验室基金(y107j6) （y107j6）
