| 抗菌肽B基因的点突变及在昆虫细胞中的表达 |
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| 引用本文： | 吴代飞 曾宪松. 抗菌肽B基因的点突变及在昆虫细胞中的表达[J]. 热带作物学报, 1999, 20(3): 54-59 |
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| 作者姓名： | 吴代飞 曾宪松 |
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| 作者单位： | 中国热带农业科学院热带作物生物技术国家重点实验室!海口５７１１０１，中国热带农业科学院热带作物生物技术国家重点实验室!海口５７１１０１，中国热带农业科学院热带作物生物技术国家重点实验室!海口５７１１０１，中国热带农业科学院热带作物生物技术国家重点实验室!海口５７ |
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| 摘 要： | 抗菌肽Ｂ基因经ＰＣＲ定点突变技术改造后，克隆到杆状病毒转移载体ｐＡｃＧＰ６７Ｂ中的信号肽位点上，随后与野生ＡｃＮＰＶ－ＤＮＡ共转染Ｓｆ９细胞，经空斑筛选、ＰＣＲ检测证实获得含该基因的重组病毒。重组病毒在Ｓｆ９细胞中扩增并表达抗菌肽Ｂ基因。取含表达产物的细胞培养液检测其生物活性表明：改造过的抗菌肽Ｂ基因在Ｓｆ９细胞中获得初步表达，且表达的分泌型产物对ＥｓｃｈｅｒｉｃｈｉａｃｏｌｉＫ１２Ｄ３１有抑菌活性。
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| 关 键 词： | 抗菌肽 PCR 杆状病毒 Sf9细胞 表达 |
Site-directed Mutagenesis of Cecropin B Geneand Expression in Insect Cells |
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| WU Daifei, ZENG Xiansong, ZHANG Yindong, PENG Cunzhi,HUANG Yadong, ZHENG Xueqin. Site-directed Mutagenesis of Cecropin B Geneand Expression in Insect Cells[J]. Chinese Journal of Tropical Crops, 1999, 20(3): 54-59 |
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| Authors: | WU Daifei ZENG Xiansong ZHANG Yindong PENG Cunzhi HUANG Yadong ZHENG Xueqin |
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| Abstract: | The Cecropin B gene, which was removed ATG codon at 5 end and added ocr-responding clone site of restriction endonuclease, was cloned into the BamH I/EcoR I siteof secretion signal sequence in baculovirus transfer vector pAcGP67B, and recombinant plasmid pAcB was produced. Cotransfection of pAcB plasmid DNA and wild type AcNPV genomic DNA into Sf9 cells and plaque purification of recombinant virus were performed afterconfirming the pAcB. A strain of recombinant virus containing the modified Cecropin Bgene was affirmed by plaque assay and PCR detection. Sf9 cells infected by the derivedrecombinant baculovirus produced and secreted antibacterial peptide B into culture fluid,showing antibacterial activity to Escherichia coli K12D31. |
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| Keywords: | antibacterial peptide PCR baculovirus Sf9 cells expression. |
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