ISOLATION AND CHARACTERIZATION OF A CLONED PORPHYROMONAS-GINGIVALIS HEMAGGLUTININ FROM AN AVIRULENT STRAIN OF SALMONELLA-TYPHIMURIUM
ISOLATION AND CHARACTERIZATION OF A CLONED PORPHYROMONAS-GINGIVALIS HEMAGGLUTININ FROM AN AVIRULENT STRAIN OF SALMONELLA-TYPHIMURIUM
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DOI：
10.1128/iai.61.3.940-946.1993
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发表时间：
1993-03-01
影响因子：
3.1
通讯作者：
BROWN, TA
中科院分区：
文献类型：
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作者：
DUSEK, DM;PROGULSKEFOX, A;BROWN, TA
牙周病致病因子牙龈卟啉单胞菌表面大分子的鉴定对研究宿主与寄生虫的相互作用以及潜在的疫苗开发具有重要意义。本研究的目的是确定克隆的牙龈假单胞菌血凝素基因能否在鼠伤寒沙门氏菌疫苗中以完整形式表达，并对重组蛋白进行鉴定。从疫苗株中纯化了重组蛋白，对其进行了鉴定，并测试了其作为竞争性血凝抑制因子的生物活性。以Luria肉汤培养的鼠伤寒沙门氏菌SL3261/pST7为研究对象，用超声破碎法对其细胞进行破碎和分级。纯化的重组蛋白能抑制牙龈假单胞菌全细胞对红细胞的凝集作用。对同一纯化蛋白的N端氨基酸序列和氨基酸组成进行了分析，发现与克隆基因的核苷酸序列预测的一致。这些结果表明，牙龈假单胞菌的表面大分子可以在沙门氏菌携带者菌株中以完整的、具有生物活性的形式表达。
Identification of surface macromolecules of Porphyromonas gingivalis that act as virulence factors in periodontal disease has important implications for studying host-parasite interactions as well as for potential vaccine development. The objective of this study was to determine whether a cloned, P. gingivalis hemagglutinin gene could be expressed in an intact form in an avirulent Salmonella typhimurium vaccine construct and to characterize the recombinant protein. The recombinant protein was purified from the vaccine strain, characterized, and tested for biological activity as a competitive inhibitor of hemagglutination. Cells of S. typhimurium SL3261/pST7 grown in Luria broth were broken by sonic disruption and fractionated. The purified recombinant protein was found to inhibit hemagglutination of erythrocytes by whole P. gingivalis cells. The same purified protein was analyzed for its N-terminal amino acid sequence and amino acid composition and found to match that predicted from the nucleotide sequence of the cloned gene. These results indicate that a surface macromolecule of P. gingivalis can be expressed in an intact and biologically active form in a Salmonella carrier strain.