中国蔬菜Issue(1)：20-25,6.
普通白菜1,5-二磷酸核酮糖羧化/加氧酶小亚基基因BcrbcS 的克隆及表达分析
Cloning and Expression Analysis of Ribulose-1,5-bisphosphate carboxylase/oxygenase Small Subunit Gene BcrbcS in Pakchoi
摘要
Abstract
The full-length cDNA sequence of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (BcrbcS)gene was cloned from the leaves of pakchoi cultivar'Suzhouqing'with resistance to downy mildew by RACE technique.This paper analyzed the expression pattern of this gene under different tissues by qRT-PCR and also prokaryotic expression characteristics of this gene by SDS-PAGE technology.Results of sequence analysis showed that the full-length cDNA of BcrbcS gene was 733 bp,among which the length of open reading frame was 543 bp.The total encodings were 181 amino acids.The molecular mass was 20.3×103 Da,and theoretical isoelectric point was 8.23.The phylogenetic analysis of amino acid homology showed that the pakchoi BcrbcS gene had similar evolutionary relationship with the other plants in the same family.Results of quantitative real-time analysis showed that the expression of BcrbcS gene was the strongest in pakchoi leaves.The expression of BcrbcS gene in pakchoi peaked at 24 hours after infection with SA and NaCl.Prokaryotic expression vector was induced by IPTG to express a fusion protein with a molecular weight of about 20×103 Da.关键词
普通白菜/1,5- 二磷酸核酮糖羧化/加氧酶小亚基基因/序列分析/qRT-PCR/原核表达Key words
Pakchoi/Ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit gene/Suquence analysis/qRT-PCR/Prokaryotic expression引用本文复制引用
刘东让,侯喜林,肖栋..普通白菜1,5-二磷酸核酮糖羧化/加氧酶小亚基基因BcrbcS 的克隆及表达分析[J].中国蔬菜,2019,(1):20-25,6.基金项目
国家重点研发计划项目(2016YFD0101701),农业部园艺作物生物学与种质创制重点实验室开放课题(IVF201804) （2016YFD0101701）
