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  gamma gene
Expression of neuronal protein synuclein gamma gene as a novel marker for breast cancer prognosis
      
These four genes are the TATA box binding protein gene, the taube nuss-like gene, the eukaryotic elongation factor 1-gamma gene, and the beta-actin-1 gene.
      
Association of interferon-gamma gene polymorphism (+874A) with arthritis manifestation in SLE
      
The human glia maturation factor-gamma gene: genomic structure and mutation analysis in gliomas with chromosome 19q loss
      
The human T-cell receptor gamma gene region spans 160 kb genomic DNA.
      
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Both tumor necrosis factor and interferon gamma Share and may work synergitically in, the activities of antivirus, antitumor and immunomodulation we fused the interferon-gamma gene and tumor necrosis factor gene in frame, with a deletion of 480 bp including translational terminator and an insertion of an artificial linking sequence in between.The fused gene was expressed in E.coli anda bif unctional hybrid protein was obtained.

肿瘤坏死因子与γ干扰素具有非常相似的生物学活性,并在一些主要的生物活性方面表现有较强的协同作用。本文采用寡核苷酸指导下的定位缺失-插入体外基因突变技术,删除了干扰素与肿瘤坏死因子串联基因之间的非编码区,去徐了γ干扰素的翻译终止信号,插入了一个人工设计的连接肽,使γ干扰素与肿瘤坏死因子基因在不改变读码框架的条件下融合起来,并在大肠杆菌表达系统中表述了兼有γ干扰素和肿瘤坏死因子功能的融合蛋白。

An E.coli DH5α strain harboring human interferon gamma gene was cultured in fermentors.The mean harvest of E.coli in three lots was 14.1g/L,and teh expression yield of the interferon was 1.02×10 9IU/L.The E.coli was homogenized and the inclusion bodies were collected.The interferon was extracted by using 7mol/L guanidine HCl.About 78.4% of bacterial proteins was removed,but the loss of biological activity of the interferon was only 10.4% in this process.The crude interferon was renatured and the biological...

An E.coli DH5α strain harboring human interferon gamma gene was cultured in fermentors.The mean harvest of E.coli in three lots was 14.1g/L,and teh expression yield of the interferon was 1.02×10 9IU/L.The E.coli was homogenized and the inclusion bodies were collected.The interferon was extracted by using 7mol/L guanidine HCl.About 78.4% of bacterial proteins was removed,but the loss of biological activity of the interferon was only 10.4% in this process.The crude interferon was renatured and the biological activity increased to 405%.The specific activity of renatured interferon also increased significantly.The mean specific activity of three lots was 1.76 ×10 7IU/mg protein.The renatured interfenon was purified by three-step chromatography.The mean recovery rate was 28.13%.The specific activities were 3.22,2.78 and 3.01×10 7IU/mg protein,respectively.The purity of three lots all was >99% determined by SDS-PAGE and >98% determined by HPLC.Other assay items were all qualified.

用带有表达质粒pBV220(含有γ干扰素基因)的大肠杆菌DH5α株进行发酵培养,3批中试的菌产量平均为14.1克/L,γ干扰素的表达量平均为1.02×109IU/L,收集的菌体经高压匀质破菌后收集包涵体,用7mol/L盐酸胍提取干扰素,此过程可去除78.4%菌体蛋白,而干扰素活性仅损失10.41%,粗制干扰素经复性可使干扰素活性提高405%,比活也有明显提高,3批平均为1.76×107IU/mg蛋白,复性γ干扰素用三步柱层析法进行纯化,其其得率平均为28.13%,其比活分别为3.22、2.78和3.01×107IU/mg蛋白。3批中试的纯化γ干扰素纯度在SDS-PAGE测定均>99%,高效液相层析(HPLC)测定>98%,其它检定项目也均合乎规程要求。

In order to improve gene expression of the prokaryotic system, it is critical to enhance the translation efficiency of mRNA. On this basis, one new expression vector pSC34 was constructed by inserting the translation enhancing sequence of T7g10L into the up stream of SD sequence within a two cistron vector pCC11 constructed also. In the same time the first cistron region of the previous vector was cut off. The properties of the vector is that it is relatively small without cI857 gene so convenient to be...

In order to improve gene expression of the prokaryotic system, it is critical to enhance the translation efficiency of mRNA. On this basis, one new expression vector pSC34 was constructed by inserting the translation enhancing sequence of T7g10L into the up stream of SD sequence within a two cistron vector pCC11 constructed also. In the same time the first cistron region of the previous vector was cut off. The properties of the vector is that it is relatively small without cI857 gene so convenient to be operated; its host strain is E.coli TAP106 with the genome containing cI857; its promoter P L is heat inducing and can be controlled stringently; the two initial ones of multiple cloning sites, Nsi Ⅰ and Sph Ⅰ, can serve for the translation initiation of target gene with or without start codon ATG respectively. By use of the vector, several genes had been overexpressed high efficiently, such as E.coli gene trxA , human protein disulfide isomerase gene, human IL 3 gene and synthetic human IFN gamma gene. It has been confirmed primarily that the vector can be used to express foreign genes with high efficiency and well generality.

1988年Olins[1]发现T7噬菌体基因10的先导序列(T7g10L)具有明显的促进基因翻译的作用.本文构建了含T7g10L的表达载体pSC34,并尝试表达了几个不同类型的基因.1材料与方法1.1菌株大肠杆菌菌株TAP106为本室储存.TAP10...

 
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