Boyle M D, Hawlitzky J, Raeder R, Podbielski A
Department of Microbiology, Medical College of Ohio, Toledo 43699-0008.
Infect Immun. 1994 Apr;62(4):1336-47. doi: 10.1128/iai.62.4.1336-1347.1994.
An emm-like gene (emmL) and a fcrA gene from group A streptococcal strain 64/14 (emmL64/14 and fcrA64/14) were amplified by PCR and force cloned into the heat-inducible expression vector pJLA 602. The emmL gene encoded a recombinant protein that bound human IgG1, IgG2, and IgG4 in a nonimmune fashion. This is the reactivity profile of a type IIa IgG-binding protein. The emmL64/14 gene product was antigenically similar to the previously identified high-molecular-weight type IIa IgG-binding protein of strain 64/14 and had an N-terminal sequence identical to that of the wild-type protein. The fcrA gene also encoded a recombinant protein with type IIa functional activity. This protein was similar to the lower-molecular-weight type IIa IgG-binding protein previously isolated from strain 64/14 and was antigenically distinct from the higher-molecular-weight type IIa protein encoded by the emmL64/14 gene. The sequences for both genes including the intervening regions are presented. The emmL gene demonstrates significant homology to other class I emm and emmL genes expressed by opacity factor-negative group A streptococcal isolates. The fcrA gene was found to be homologous to other fcrA genes normally present in opacity factor-positive group A isolates. The sequence upstream of the fcrA gene and the intervening sequence between the end of the fcrA gene and the start of the emmL gene were similar to those reported for other fcrA genes.
通过聚合酶链反应(PCR)扩增A组链球菌菌株64/14中的一个类emm基因(emmL)和一个fcrA基因(emmL64/14和fcrA64/14),并强制克隆到热诱导表达载体pJLA 602中。emmL基因编码一种重组蛋白,该蛋白以非免疫方式结合人IgG1、IgG2和IgG4。这是IIa型IgG结合蛋白的反应特性。emmL64/14基因产物在抗原性上与先前鉴定的64/14菌株的高分子量IIa型IgG结合蛋白相似,并且其N端序列与野生型蛋白相同。fcrA基因也编码一种具有IIa型功能活性的重组蛋白。该蛋白与先前从64/14菌株中分离出的低分子量IIa型IgG结合蛋白相似,并且在抗原性上与emmL64/14基因编码的高分子量IIa型蛋白不同。给出了包括间隔区在内的两个基因的序列。emmL基因与不透明因子阴性A组链球菌分离株表达的其他I类emm和emmL基因具有显著同源性。发现fcrA基因与通常存在于不透明因子阳性A组分离株中的其他fcrA基因同源。fcrA基因上游的序列以及fcrA基因末端与emmL基因起始之间的间隔序列与其他fcrA基因报道的序列相似。