Cloning of the structural genes of three H8 antigens and of protein III of Neisseria gonorrhoeae.

Abstract

A bank of gonococcal DNA was constructed in the .lambda. gt11 expression vector. Immunological screening of the bank resulted in the isolation of a clone that contains the structural gene of protein III. In addition, several clones reactive with mAbs specific for the H8 antigen were isolated. DNA hybridization studies revealed that these H8-reactive clones were derived from three different gonococcal genes. When the products produced by these clones were used to absorb antibodies from a rabbit antiserum, and the eluted antibodies were used in immunological studies, it could be shown that the parent gonococcus expressed the product of two of these H18 genes, and in strain R10, these had Mr of .apprx. 19,700 21,200 respectively. The larger form has not been recognized hitherto because the epitope reactive with the H8 mAb may be masked in this product. PIII. We are equally indebted to Dr. Penny Hitchcock for giving us the mAb to the H8 antigen and sharing her potocols for Western blotting of this antigen.