Expression by Streptomyces lividans of the Rat α Integrin CD11b A-Domain as a Secreted and Soluble Recombinant Protein

• Published in: Journal of biomedicine & biotechnology Vol. 2007; pp. 1 - 6
• Main Authors: Ayadi, Dorra Zouari, Chouayekh, Hichem, Mhiri, Sonda, Zerria, Khaled, Fathallah, Dahmani M, Bejar, Samir
• Format: Journal Article
• Language: English
• Published: Hindawi Limited 01.01.2007; Hindawi Publishing Corporation
• Subjects: Biotechnological microorganisms; Cardiology and cardiovascular system; Cellular Biology; Gene expression; Genetic aspects; Human health and pathology; Integrins; Life Sciences; Microbial biotechnology; Streptomyces; Tunisia; Valve disorders; Genetics; Congenital heart disease; Cardiomyopathy; Non-compaction; Neurologic involvement
• ISSN: 1110-7243; 1110-7251
• DOI: 10.1155/2007/54327

We already reported the use of a long synthetic signal peptide (LSSP) to secrete the Streptomyces sp. TO1 amylase by Streptomyces lividans strain. We herein report the expression and secretion of the rat CD11b A-domain using the same LSSP and S. lividans as host strain. We have used the Escherichia coli/Streptomyces shuttle vector pIJ699 for the cloning of the A-domain DNA sequence downstream of LSSP and under the control of the constitutive erm E- up promoter of Streptomyces erythraeus . Using this construct and S. lividans as a host strain, we achieved the expression of 8 mg/L of soluble secreted recombinant form of the A-domain of the rat leukocyte β 2 integrin CD11/CD18 alpha M subunit (CD11b). This secreted recombinant CD11b A-domain reacted with a function blocking antibody showing that this protein is properly folded and probably functional. These data support the capability of Streptomyces to produce heterologous recombinant proteins as soluble secreted form using the “LSSP” synthetic signal peptide.