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| Large Protein Production Through Split Intein-Mediated Trans-Splicing |
| ZHANG Jing,LIU Huan,ZHOU Jing,LIU Jian-hua |
| College of Life Science & Biotechnology, Shanghai Jiao Tong University, Shanghai 200240,China |
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Abstract Npu DnaE intein was used to produce some large proteins, which were difficult to obtain through conventional expression systems. A T7 expression system was described, by which the gene of T7 RNA polymerase is split into two pieces, and each piece fuses with Npu DnaE N and C terminal sequences respectively. Functional T7 RNA polymerase is created by mixing the two kinds of fusion constructs in vitro.The approach of split inteinmediated production of large proteins, in theory, readily generalizable to the purification of other large, cytotoxic or membrane proteins.
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Received: 09 September 2009
Published: 21 December 2009
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