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1.
公开(公告)号:US20110165583A1
公开(公告)日:2011-07-07
申请号:US13061640
申请日:2009-09-02
摘要: A modified vector comprising a reporter gene having a STOP codon upstream of the multiple cloning site of the vector which is characterized in that the recombinant clones show fluoresce or show color in presence of inducer. A method for identification and selection of recombinant clones comprising the modified vector wherein the recombinant clones florescence or show color in a suitable suppressor strain of the STOP codon associated with the gene of interest. A method of preparation of recombinant clone comprising gene of interest and modified vector comprising amplification of gene of interest using specific primers containing STOP codon different from STOP codon used with reporter gene; cloning the amplified gene of interest in the modified vector; transformation of cloned modified vector in the STOP codon suppressor host cell wherein the STOP codon suppressor host cell is specific for STOP codon used with the gene of interest wherein the recombinant clones either fluorescence or show color depending upon the reporter gene used.
摘要翻译: 包含在载体的多克隆位点上游具有一个末端密码子的报道基因的修饰载体,其特征在于重组克隆在诱导物存在下显示出荧光或显示出颜色。 用于鉴定和选择包含修饰载体的重组克隆的方法,其中所述重组克隆在与所关心的基因相关的STOP密码子的合适的抑制菌株中荧光或显示颜色。 包含目的基因和修饰载体的重组克隆的制备方法,其包括使用与报告基因使用的不同于STOP密码子的不同密码子的特异性引物扩增目标基因; 在经修饰的载体中克隆所扩增的目的基因; 克隆的修饰载体在STOP密码子抑制基因宿主细胞中的转化,其中所述的停止密码子抑制基因宿主细胞对于与感兴趣的基因一起使用的终止密码子是特异性的,其中所述重组克隆根据所使用的报道基因是荧光或显示颜色。
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公开(公告)号:US08853380B2
公开(公告)日:2014-10-07
申请号:US13702028
申请日:2011-06-03
申请人: Shardul Salunkhe , Bhaskarjyoti Prasad , Praveen Muneshwar , Anjali Apte Deshpande , Sriram Padmanabhan
发明人: Shardul Salunkhe , Bhaskarjyoti Prasad , Praveen Muneshwar , Anjali Apte Deshpande , Sriram Padmanabhan
IPC分类号: C07H21/04 , C12N9/00 , C12P21/06 , C07K14/555 , C07K14/535 , C07K14/54 , C07K14/49 , C07K14/55 , C12N9/96 , C07K14/635 , C12N9/72 , C07K14/31 , C07K14/485
CPC分类号: C07K14/635 , C07K14/31 , C07K14/485 , C07K14/49 , C07K14/535 , C07K14/5403 , C07K14/5406 , C07K14/5409 , C07K14/5412 , C07K14/5431 , C07K14/55 , C07K14/555 , C07K2319/35 , C07K2319/50 , C12N9/6459 , C12N9/96 , C12P21/06 , C12Y304/21069
摘要: The present invention relates to modified SAK gene having amino acid SEQ ID 2. The present invention further relates to process for cloning and expressing modified SAK gene fusion protein which imparts improved stability to the heterologous protein of interest. Further the invention relates to process of purification of recombinant heterologous proteins from bacterial inclusion bodies using modified SAK.
摘要翻译: 本发明涉及具有氨基酸SEQ ID No.2的经修饰的SAK基因。本发明还涉及克隆和表达修饰的SAK基因融合蛋白的方法,其赋予所感兴趣的异源蛋白质更好的稳定性。 此外,本发明涉及使用改良的SAK从细菌包涵体纯化重组异源蛋白质的方法。
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公开(公告)号:US20130122542A1
公开(公告)日:2013-05-16
申请号:US13702028
申请日:2011-06-03
申请人: Shardul Salunkhe , Bhaskarjyoti Prasad , Praveen Muneshwar , Anjali Apte Deshpande , Sriram Padmanabhan
发明人: Shardul Salunkhe , Bhaskarjyoti Prasad , Praveen Muneshwar , Anjali Apte Deshpande , Sriram Padmanabhan
CPC分类号: C07K14/635 , C07K14/31 , C07K14/485 , C07K14/49 , C07K14/535 , C07K14/5403 , C07K14/5406 , C07K14/5409 , C07K14/5412 , C07K14/5431 , C07K14/55 , C07K14/555 , C07K2319/35 , C07K2319/50 , C12N9/6459 , C12N9/96 , C12P21/06 , C12Y304/21069
摘要: The present invention relates to modified SAK gene having amino acid SEQ ID 2. The present invention further relates to process for cloning and expressing modified SAK gene fusion protein which imparts improved stability to the heterologous protein of interest. Further the invention relates to process of purification of recombinant heterologous proteins from bacterial inclusion bodies using modified SAK.
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