Gene splicing method and products produced therefrom

Chemistry: molecular biology and microbiology – Treatment of micro-organisms or enzymes with electrical or... – Modification of viruses

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435 68, 435 91, 435253, 435317, 435874, 435875, 435 23, 435 29, 435 56, 435 72, 435 80, C12N 1500, C12N 120, C12N 100, C12P 2100, C12P 1934, C12R 138, C12R 1385

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045088233

ABSTRACT:
An improved gene splicing and recombinant plasmid transformation method is described. The method includes mechanical fragmenting of chromosomal DNA followed by conventional digestion with a restriction enzyme and gene splicing into a vector to provide recombinant plasmids in a bank of at least about 100 different plasmids. The plasmids in the bank are provided for transformation into a suitable host, particularly a plasmid free bacterium of the same species from which the chromosomal DNA or the vector is derived. The method provides high transformation frequencies because of the presence of multiple "super coiled" or closed coiled recombinant plasmids in the bank. The method also allows for the direct selection of many different phenotypic traits in a pool of the transformed hosts. The selected hosts are useful for the production of various gene products.

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