Method for enhancing solubility of recombinant protein...

Chemistry: molecular biology and microbiology – Vector – per se

Reexamination Certificate

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Reexamination Certificate

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07056732

ABSTRACT:
The present invention relates to the field of biotechnology. The invention provides a novel approach using tobacco mosaic virus omega leader sequence to enhance the solubility of the recombinant products inE. coliand the method of use therefore. The invention provides the utilization of tobacco mosaic virus omega leader sequence intoE. coliexpression vector, and the tobacco mosaic virus omega leader sequence containing expression vector can be used in combination with other available means to obtain higher expression or better solubility. The invention can be applied to biotechnological pharmaceutical industry, genetic engineering, biochemistry and molecular biology etc. The invention provides an expression vector pTORG, which is a highly efficient GST fusion expression vector, and can significantly enhance the yield of biologically active recombinant products.

REFERENCES:
Gallie et al. The 5′-leader se1uence of tobacco masaic virus RNA enhances the expression of foreign gene transcripts in vitro and in vivo Nucleic Acids Research vol. 15 No. 8 1987 pp. 3257-3273.
Ausubel et al. (Short Protocols in Molecular Biology 1999 pp. 10-76 to 10-80 and 16-24 to 16-27.
“Nucleotide Sequence at the 5′ Extremity of Tobacco-Mosaic-Virus RNA”, Richards et wl., Eur. J. Biochem. 84, 513-519 (1978).
“Identification of the motifs within the tobacco mosaic virus 5′-leader responsible for enhancing translation”, Gallie et al., Nucleic Acids Research, vol. 20, No. 17, 4631-4638.
“pALEX, a dual-tag prokaryotic expression vector for the purification of full-length proteins”, Panagiotidis et al., Gene, 164 91995) 45-47.

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