The cloned bacteriophage T4 pin gene functions to stabilize several different kinds of proteins in Escherichia coli bacteria. Incomplete proteins such as puromycyl polypeptides, abnormal but complete proteins such as the A phage tsO protein, and labile eukaryotic proteins encoded by genes cloned in E. coli such as mature human fibroblast interferon are stabilized in cells in which the T4 pin gene is expressed. The cloned T4 pin gene does not seem to affect the turnover of normal E. coli proteins.The introduction of cloned genes into Escherichia coli bacteria for the purpose of producing large amounts of specific proteins not normally present in E. coli cells has led to an examination of various problems associated with the expression of such cloned genes. Difficulties in obtaining expected levels of expression from genes cloned in bacterial cells may be encountered at the levels of stability of the gene and its vector within the host cell (1, 2), transcription of the cloned gene (3-7), and translation of its mRNA into protein (5,6,8 Difco agar, maltose/L bottom agar is identical to maltose/L top agar. Bacteria that were to be assayed for interferon activity were grown in the presence of the appropriate antibiotic(s) in either M9, which contains, per liter: NH4Cl, 1.0 g; KH2PO4, 3.0 g; Na2HPO4, 6.0 g; NaCl, 5.0 g; MgSO4 7H2O, 0.25 g; CaCl2, 0.015 g; glucose, 2.0 g; supplemented with vitamin BI and Casamino acids to final concentrations of 0.001% and 0.5%, respectively, 2059The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.
Advantages of lambda as a cloning vector are discussed along with considerations for the insert DNA (i.e., size, spi and hfl state). Maps of lambda-derived cloning vectors are provided in addition to a discussion and map of a cosmid (a lambda-derived plasmid vector).
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