A structural gene with 385 bp, which codes for the mature small subunit (rbcS) of ribulose-1,5-bi9phosphate carboxylase/oxygenase from tobacco, has been redesigned according to the codon usage of E.coli and chemically...A structural gene with 385 bp, which codes for the mature small subunit (rbcS) of ribulose-1,5-bi9phosphate carboxylase/oxygenase from tobacco, has been redesigned according to the codon usage of E.coli and chemically synthesized. To facilitate the systematic investigation of structure-functional relationship of the rbcS by site-specific mutagenesis, twenty one unique restriction sites distributed throughout the synthetic gene were designed. Gene synthesis was started from chemically synthesizing sixteen oligonucleotides each with 23-66 nucleotides, and these oligonu-cleotides were annealed to form eight duplexes from which 5'- and 3'- two half molecules were formed by stepwise T4 DNA ligase reaction, and then each half molecule was cloned into plasmid pWR13, after that, two half molecules were further confirmed by DNA sequencing, finally both half molecules excised from the cloning plasmid were recombinated to form plasmid pCOTrbcS containing the whole structural gene for tobacco rbcS.展开更多
自上世纪80年代核酮糖-1,5-二磷酸羧化酶/加氧酶活化酶(Ru Bis CO activase,RCA)被发现后,一直广受关注,其酶学特性、体内外活化及其对光合作用的影响、体内外活性调节机制、基因和分子结构、反义突变和超表达等研究,已成为了光合研究...自上世纪80年代核酮糖-1,5-二磷酸羧化酶/加氧酶活化酶(Ru Bis CO activase,RCA)被发现后,一直广受关注,其酶学特性、体内外活化及其对光合作用的影响、体内外活性调节机制、基因和分子结构、反义突变和超表达等研究,已成为了光合研究领域的一大热点。本文结合国内外研究进展,就RCA分子结构、与光合作用的关系,特别是其在植物抗逆中的作用等方面进行综述。展开更多
基金Project supported by grants from the High Technology Development Program of China,From the National Key Laboratory of Biooganic and Natural Product Chemistry of China,and from the Laboratory of Computer Chemistry,Chinese Academy of Sciences.
文摘A structural gene with 385 bp, which codes for the mature small subunit (rbcS) of ribulose-1,5-bi9phosphate carboxylase/oxygenase from tobacco, has been redesigned according to the codon usage of E.coli and chemically synthesized. To facilitate the systematic investigation of structure-functional relationship of the rbcS by site-specific mutagenesis, twenty one unique restriction sites distributed throughout the synthetic gene were designed. Gene synthesis was started from chemically synthesizing sixteen oligonucleotides each with 23-66 nucleotides, and these oligonu-cleotides were annealed to form eight duplexes from which 5'- and 3'- two half molecules were formed by stepwise T4 DNA ligase reaction, and then each half molecule was cloned into plasmid pWR13, after that, two half molecules were further confirmed by DNA sequencing, finally both half molecules excised from the cloning plasmid were recombinated to form plasmid pCOTrbcS containing the whole structural gene for tobacco rbcS.
文摘自上世纪80年代核酮糖-1,5-二磷酸羧化酶/加氧酶活化酶(Ru Bis CO activase,RCA)被发现后,一直广受关注,其酶学特性、体内外活化及其对光合作用的影响、体内外活性调节机制、基因和分子结构、反义突变和超表达等研究,已成为了光合研究领域的一大热点。本文结合国内外研究进展,就RCA分子结构、与光合作用的关系,特别是其在植物抗逆中的作用等方面进行综述。
