Expression of tuna growth hormone cDNA in Escherichia coli

In order to produce tuna (Thunnus thynnus ) growth hormone (GH), expression plasmid (pUES13S) carrying tuna GH cDNA was constructed using a vector (pKK223-3), in which the replication origin was replaced with that of pUC19. The expression of the tuna GH cDNA was greatly affected by the distance betw...

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Veröffentlicht in:	Applied microbiology and biotechnology 1989-02, Vol.30 (2), p.153-159
Hauptverfasser:	SATO, N, HAYAMI, T, MURATA, K, WATANABE, K, KARIYA, Y, SAKAGUCHI, M, KIMURA, S, NONAKA, M, KIMURA, A
Format:	Artikel
Sprache:	eng
Schlagworte:	Applied microbiology Biological and medical sciences Biotechnology cDNA DNA Escherichia coli Fundamental and applied biological sciences. Psychology gene expression gene transfer Genetic engineering Genetic technics growth hormone growth regulators hormones Marine Methods. Procedures. Technologies Microbiology Miscellaneous plasmids Thunnus thynnus Vectors (cloning, transfer, expression). Insertion sequences and transposons
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Zusammenfassung:	In order to produce tuna (Thunnus thynnus ) growth hormone (GH), expression plasmid (pUES13S) carrying tuna GH cDNA was constructed using a vector (pKK223-3), in which the replication origin was replaced with that of pUC19. The expression of the tuna GH cDNA was greatly affected by the distance between a Shine-Dalgarno (SD) sequence and the initiation condon (ATG) and was most efficient when the distance was adjusted to 13 base pairs (bp). The amount of tuna GH produced by Escherichia coli JM109 with pUES13S was more than 12.5% of the total cytosolic proteins and the product was immunologically identified to be tuna GH (mol. wt. 21,000) by Western blot analysis using tuna GH specific immunoglobulin G (IgG).
ISSN:	0175-7598 1432-0614
DOI:	10.1007/bf00264004