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Preliminary Study on the Expression of BGT gene in Transgenic Birch

机译:BGT基因在转基因桦树中表达的初步研究

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In this paper, Southern blot analysis of the transgenic birch (Betula platyphylla Suk.) plants indicated that the the copy number of bgt varies from one to four. Of which 18.2 percent were single copy. Transcriptional expression of bgt gene in transgenic birch plants was analyzed by Northern blot. TGS has been found in four transgenic lines with one, three and four copies of bgt. Bgt gene was inserted into the prokaryotic vector pET 28a and transferred into E.coli BL21 (DE3) for expression of BGT fusion protein. After overnight cleavage with Thrombin and purification by immobilized metal ion affinity chromatography, the final BGT proteins were obtained with purity of 90%. The ELISA result showed that the BGT protein expression level in the transgenic birch plants ranged from 0.000% to 0.283% of total soluble protein. No significant correlation was found between copy number and expression level of bgt gene.
机译:在本文中,对转基因桦(Betula platyphylla Suk。)植物的Southern印迹分析表明,bgt的拷贝数从一到四不等。其中18.2%是单拷贝。通过Northern印迹分析bgt基因在转基因桦树植物中的转录表达。已经在具有一,三和四个拷贝的bgt的四个转基因品系中发现了TGS。将Bgt基因插入原核载体pET 28a中,并转移至大肠杆菌BL21(DE3)中以表达BGT融合蛋白。用凝血酶裂解过夜并通过固定的金属离子亲和层析纯化后,获得最终的BGT蛋白,纯度为90%。 ELISA结果表明,转基因桦树植物中BGT蛋白的表达量占可溶性蛋白总量的0.000%至0.283%。在bgt基因的拷贝数和表达水平之间没有发现显着的相关性。

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