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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Construction of two gateway vectors for gene expression in fungi.
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Construction of two gateway vectors for gene expression in fungi.

机译：用于在真菌中表达基因的两个通道载体的构建。

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摘要

We report the construction of two Gateway fungal expression vectors pCBGW and pGWBF. The pCBGW was generated by introducing an expression cassette, which consists of a Gateway recombinant cassette (attR1-Cmr-ccdB-attR2) under the control of fungal promoter PgpdA and a terminator TtrpC, into the multiple cloning site of fungal vector pCB1004. The pGWBF is a binary vector, which was generated from the plant expression vector pGWB2 by replacing the CaMV35S promoter with PgpdA. The pGWBF can be transformed into fungi efficiently with Agrobacterium-mediated transformation. The applicability of two newly constructed vectors was tested by generating the destination vectors pGWBF-GFP and pCBGW-GFP and examining the expression of GFP gene in Trichoderma viride and Gibberella fujikuroi, respectively. Combining with the advantage of Gateway cloning technology, pCBGW and pGWBF will be useful in fungi for large-scale investigation of gene functions by constructing the interested gene destination/expression vectors in a high-throughput way.
机译：我们报告了两个网关真菌表达载体pCBGW和pGWBF的建设。通过将表达盒引入到真菌载体pCB1004的多克隆位点来表达pCBGW，该表达盒由在真菌启动子PgpdA和终止子TtrpC的控制下的Gateway重组盒（attR1-Cmr-ccdB-attR2）组成。 pGWBF是二元载体，其是通过用PgpdA替换CaMV35S启动子从植物表达载体pGWB2产生的。用农杆菌介导的转化可以将pGWBF有效地转化为真菌。通过生成目的载体pGWBF-GFP和pCBGW-GFP并检查GFP基因在绿色木霉和藤本赤霉菌中的表达，测试了两个新构建的载体的适用性。结合网关克隆技术的优势，pCBGW和pGWBF将通过以高通量的方式构建感兴趣的基因目的地/表达载体，在真菌中用于基因功能的大规模研究。

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