Acyl carrier protein-I/protein-A gene fusion, products and metho

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...

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536 27, 4351723, 4353201, 530370, C12P 2102, C12N 1562, C07K 300

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active

050875639

ABSTRACT:
A gene encoding spinach acyl carrier protein-I-Gly.sup.76 (ACP-I) was fused to a gene encoding the F.sub.c -binding portion of staphylococcal protein A. The gene fusion can be expressed in E. coli thereby resulting in the production of an appropriately sized protein possessing ACP activity in acyl ACP synthetase reactions, plant MCT reactions, and plant fatty acid synthesis. This gene fusion, under the control of the .lambda.P.sub.R promoter, expresses high levels of a 42 kDa fusion protein in E. coli. This protein functions as a affinity moiety in purification methods for the isolation of enzymes involved in fatty acid synthetase, acyl ACP synthetase and malonyl-CoA:ACP tranacylase reactions.

REFERENCES:
Arch. Biochem. Biophys. 256:90-100, Jul. 1987, Beremand et al., Synthesis, Cloning and Expression in Eschericlaia Coli of a Spinach Acyl Carrier Protein-I Gene.
EMBO J. 4:1075-1080, 1984, Nilsson et al. Immobilization and Purification of Enzymes with Staphylococcal Protein A gene fusion Vectors.
John B. Ohlrogge et al., "Acyl Carrier Protein: Central Cofactor for Plant Fatty Acid Metabolism," What's New in Plant Physiology 15(11): 41-44 (Nov. 1984).
Katherine M. Schmid et al., "A Root Acyl Carrier Protein-II from Spinach is Also Expressed in Leaves and Seeds," Plant Mol. Biol. 15: 765-778 (1990).
John B. Ohlrogge, Table: "ACP% Protein Identity," personal communication to Phillip D. Beremand.

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