| Cloning and Sequencing of Trichosanthin Gene and Its Expression in Escherichia coli and Tobacco Plant |
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| 引用本文： | 鲍一明,储瑞银,韩晋华,张会,潘乃穟,顾孝诚,陈章良. Cloning and Sequencing of Trichosanthin Gene and Its Expression in Escherichia coli and Tobacco Plant[J]. 中国科学B辑(英文版), 1993, 0(6) |
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| 作者姓名： | 鲍一明 储瑞银 韩晋华 张会 潘乃穟 顾孝诚 陈章良 |
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| 作者单位： | The National Laboratory of Protein Engineering and Plant Genetic Engineering，Department of Biology，Peking University，Beijing 100871，PRC，The National Laboratory of Protein Engineering and Plant Genetic Engineering，Department of Biology，Peking University，Beijing 100871，PRC，The National Laboratory of Protein Engineering and Plant Genetic Engineering，Department of Biology，Peking University，Beijing 100871，PRC，The National Laboratory of Protein Engineering and Plant Genetic Engineering，Department of Biology，Peking University，Beijing 100871，PRC，The National Laboratory of Protein Engineering and Plant Genetic Engineering，Department of Biology，Peking University，Beijing 100871，PRC，The National Laboratory of Protein Engineering and Plant Genetic Engineering，Department of Biology，Peking University，Beijing 100871，PRC，The National Laboratory of Protein Engineering and Plant Genetic Engineering，Department of Biology，Peking University，Beijing 100871，PRC |
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| 摘 要： | A Trichosanthin gene was cloned from Trichosanthes kirilowii genomic DNA by polymerase chain reaction (PCR). Nucleotide sequence data indicated that we obtained the coding region of the mature Trichosanthin peptide as well as its signal peptide at the N-terminus. Comparisons of our sequence with the previously reported nucleotide sequences of this gene showed 99.25% homology, yet there were notable differences between the previously reported amino acid sequence and our deduced result. This gene was subcloned into a highlevel expression plasmid (pJLA502) of E. coli under the control of a P_RP_L promoter, and we observed the gene product after temperature induction. The gene was further cloned into plant intermediate vector pE3 under the control of a CaMV 35S promoter, and transferred into a tobacco genome using the agrobacterium-mediated gene transfer system. Western blotting analysis of the protein extracted from Escherichia coli and transgenic tobacco plants proved that the Trichosanthin gene has been
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Cloning and Sequencing of Trichosanthin Gene and Its Expression in Escherichia coli and Tobacco Plant |
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| BAO Yi-Ming CHU Rui-Yin HAN Jin-HuaZHANG Hui PAN Nai-Sui GU Xiao-Cheng CHEN Zhang-Liang. Cloning and Sequencing of Trichosanthin Gene and Its Expression in Escherichia coli and Tobacco Plant[J]. Science in China(Chemistry), 1993, 0(6) |
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| Authors: | BAO Yi-Ming CHU Rui-Yin HAN Jin-HuaZHANG Hui PAN Nai-Sui GU Xiao-Cheng CHEN Zhang-Liang |
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| Abstract: | A Trichosanthin gene was cloned from Trichosanthes kirilowii genomic DNA by polymerase chain reaction (PCR). Nucleotide sequence data indicated that we obtained the coding region of the mature Trichosanthin peptide as well as its signal peptide at the N-terminus. Comparisons of our sequence with the previously reported nucleotide sequences of this gene showed 99.25% homology, yet there were notable differences between the previously reported amino acid sequence and our deduced result. This gene was subcloned into a highlevel expression plasmid (pJLA502) of E. coli under the control of a P_RP_L promoter, and we observed the gene product after temperature induction. The gene was further cloned into plant intermediate vector pE3 under the control of a CaMV 35S promoter, and transferred into a tobacco genome using the agrobacterium-mediated gene transfer system. Western blotting analysis of the protein extracted from Escherichia coli and transgenic tobacco plants proved that the Trichosanthin gene has been expressed in both organisms. |
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| Keywords: | Trichosanthin polymerase chain reaction nucleotide sequence analysis gene expression transgenic plant |
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