IN VITRO SYNTHESIS OF B. CEREUS T SPORE COAT PROTEIN

WILLIE WRIGHT, Purdue University

Abstract

In vitro systems have been developed to investigate questions concerning the regulation of the synthesis of the spore coat protein of B. Cereus T. The results of these studies indicate that the spore coat protein is synthesized in relatively large amounts beginning at the second hour after the end of exponential growth, with lower levels of synthesis at the end of exponential growth and some indication of a lower level of a lower level of synthesis with RNA extracted from exponentially growing cells. This time of maximal synthesis correlates well with the onset of synthesis of the protein in vivo, as determined by antibody precipitation data. In vitro translation studies also confirm the synthesis of the spore coat protein as a precursor of 65 Kd which is also consistent with in vivo studies. There is also some evidence of a polypeptide of 90 Kd reacting with spore coat antibody. Although a direct assessment of the halflife of the mRNA responsible for the synthesis of the spore coat precursor was not possible, two types of analyses indicate a greater stability than other messengers present in the cell at the same time. When the synthesis of the coat precursor was examined in E. coli, exponential B. cereus and postexponential B. cereus in vitro systems, the postexponential extracts proved to be most efficient, while the E. coli was the least efficient, and the exponential B. Cereus extract was intermediate. In addition to the synthesis of the spore coat protein, the three in vitro systems are capable of synthesizing glutamine synthetase monomers directed by RNA extracted from exponential B. subtilis cells.

Degree

Ph.D.

Subject Area

Biology

COinS