| 黑曲霉木聚糖酶基因（xynB）的克隆及真核分泌表达 |
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| 引用本文： | 张茂,周庆丰,杜云平,刘德武,孟繁明,周美华,吴珍芳.黑曲霉木聚糖酶基因（xynB）的克隆及真核分泌表达[J].农业生物技术学报,2010,18(2):368-373. |
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| 作者姓名： | 张茂 周庆丰 杜云平 刘德武 孟繁明 周美华 吴珍芳 |
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| 作者单位： | 华南农业大学动物科学学院,广州,510642;广东省温氏集团研究院,新兴,527400 |
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| 基金项目： | 国家高技术研究发展计划(863计划)，国家重大科技专项项目 |
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| 摘 要： | 通过RT-PCR方法,以黑曲霉(Aspergillus niger)GIM3.452总RNA为模板,克隆出木聚糖酶B(xy-lanase B,xynB)基因的成熟肽编码序列(567 bp),编码188个氨基酸.将其与猪腮腺分泌蛋白(parotid secretoryprotein,PSP)基因的信号肽序列通过重叠延伸PCR(SOE-PCR)得到拼接片段PSxynB,并将其克隆到真核表达载体pcDNA6/HisTMA中,得到重组质粒pcDNA-PSxynB,重组质粒经过酶切、测序鉴定,证实含有目的片段,且构建正确.在脂质体介导下将重组质粒pcDNA-PSxynB转染猪肾细胞(PK15),通过RT-PCR证实其在PK15细胞中表达,并在细胞培养液中测到了木聚糖酶活最高达36.4 IU/mL.
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| 关 键 词： | 黑曲霉 木聚糖酶 分泌表达 |
Cloning and Eukaryotic Secrete Expressing Xylanase Gene(xynB) from Aspergillus niger |
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| Zhang Mao,Zhou Qingfeng,Du Yunping,Liu Dewu,Meng Fanming,Zhou Meihua,Wu Zhenfang.Cloning and Eukaryotic Secrete Expressing Xylanase Gene(xynB) from Aspergillus niger[J].Journal of Agricultural Biotechnology,2010,18(2):368-373. |
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| Authors: | Zhang Mao Zhou Qingfeng Du Yunping Liu Dewu Meng Fanming Zhou Meihua Wu Zhenfang |
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| Institution: | Zhang Mao Zhou Qingfeng Du Yunping Liu Dewu Meng Fanming Zhou Meihua Wu Zhenfang College of Animal Science,South China Agricultural University,Guangzhou,510642,Institute of Wen's Group,Guangdong Province,Xinxing,527400 |
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| Abstract: | The mature peptide coding sequence of xylanase gene(xynB) was amplified by RT-PCR from Aspergillus niger GIM3.452 total RNA extracts.The result showed that the mature peptide sequence of xynB was consisted of 567 bp, and encoded 188 amino acids.Then, the mature peptide sequence of xynB gene and signal peptide sequence of pig parotid secretory protein gene(PSP) were spliced by overlap extension PCR(SOE-PCR), which PSxynB was obtained.PSxynB was subcloned into the eukaryotic expressing plasmid vector pcDNA6/H... |
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| Keywords: | Aspergillus niger Xylanase Secrete expression |
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