Variants of Neisseria gonorrhoeae MS11 show distinct colony morphologies because of the expression of a class of surface components called opacity (Opa, PII) proteins. Southern analyses combined with molecular cloning of genomic DNA from a single variant of MS11 has identified 11 opa genes contained in separate loci. These opa genes code for distinct opacity proteins which are distinguishable at their variable domains. The opa gene analyses were also extended to divergent variants of MS11. These studies have shown that, during in vitro and in vivo culture, 10 of the 11 opa genes did not undergo significant change in their primary sequence. However, in these variants, one gene (opaE) underwent non-reciprocal inter-opa recombinations to generate newer Opa variants. Phylogenic analysis of the opa gene sequences suggests that the opa gene family have evolved by a combination of gene duplication, gene replacement and partial inter-opa recombination events.
Attachment of Neisseria gonorrhoeae to amnion cells in tissue culture is facilitated if the gonococci bear pili. This has been determined by studying the number of pilated, colony type 2 gonococci associated with amnion cells after incubation in vitro as compared with the number of nonpilated, colony type 4 gonococci present with amnion cells under the same conditions. These data are supported by light microscope findings. Electron microscope studies provide visualization of fine structure of gonococcal attachment. Gonococci are also found within amnion cells in this in vitro system.
When pilus+ Gc were introduced into a male subject's urethra, they gave rise to pilus+ variants whose pilin mRNAs differed from that of input Gc. The differences stemmed from the Gc genome's single complete pilin gene having undergone gene conversion by different partial pilin genes' sequences and by different length stretches of a single partial pilin gene. In some instances, the variant's pilin mRNA appeared to reflect two independent gene-conversion events that used sequences from two different partial pilin genes. The resulting variants' pilins exhibited antigenic differences compared with the pilin polypeptide of input Gc; these differences were discernible by immunoblotting with mAbs. Amino acid and antigenic changes occurred in a segment of the variants' pilin polypeptides that previously was thought to be conserved or constant in sequence.
Colonies with markedly differing color and opacity characteristics were found for Neisseria gonorrhoeae propagated on translucent, solid medium. These optical properties of gonococcal colonies were independent of pilation and appeared to be related to the degree of aggregation among the gonococci comprising the colonies. Dark, opaque colonies contained highly aggregated gonococci which were more susceptible to killing and to solubilization of their 125-labeled surface components than were organisms comprising light, transparent colony forms.
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