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[蜘蛛丝体壁蛛丝蛋白1类似物的合成基因构建及其在烟草植株中的表达]

[Construction of the synthetic genes for protein analogs of spider silk carcass spidroin 1 and their expression in tobacco plants].

作者信息

Piruzian E S, Bogush V G, Sidoruk K V, Goldenkova I V, Musiĭchuk K A, Debabov V G

机构信息

Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, 119991 Russia.

出版信息

Mol Biol (Mosk). 2003 Jul-Aug;37(4):654-62.

Abstract

To obtain transgenic tobacco plants expressing recombinant analogs of spider dragline silk spidroin 1, artificial 1f5 and 1f9 coding for spidroin 1 analogs were 3'-fused in-frame with the reporter lichenase gene. The Tr2' weak constitutive promoter of Agrobacterium tumefaciens T-DNA and the strong constitutive promoter of the cauliflower mosaic virus 35S RNA gene were used as regulatory elements. The expression cassettes were used to transform agrobacteria and then introduced in tobacco leaf disks. On evidence of Southern hybridization, transgenic plants each carried a single copy of a hybrid gene, which corresponded in size to the constructed one. Zymography and Western blotting revealed full-length hybrid proteins in leaf extracts of transgenic plants. The results testified that plants can maintain and express synthetic genes for spider silks and, consequently, may be used as a convenient producer of recombinant silk analogs.

摘要

为了获得表达蜘蛛拖牵丝蛛丝蛋白1重组类似物的转基因烟草植株,将编码蛛丝蛋白1类似物的人工1f5和1f9在框架内与报告基因地衣酶基因进行3'融合。根癌农杆菌T-DNA的Tr2'弱组成型启动子和花椰菜花叶病毒35S RNA基因的强组成型启动子用作调控元件。将表达盒用于转化农杆菌,然后导入烟草叶盘。根据Southern杂交的证据,转基因植物各自携带一个杂交基因的单拷贝,其大小与构建的基因相对应。酶谱分析和蛋白质免疫印迹法揭示了转基因植物叶片提取物中的全长杂交蛋白。结果证明植物可以维持并表达蜘蛛丝的合成基因,因此可以用作重组丝类似物的便利生产者。

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