DNA-SEQUENCE DETERMINATION AND BIOCHEMICAL-ANALYSIS OF THE IMMUNOGENIC PROTEIN P36, THE LACTATE-DEHYDROGENASE (LDH) OF MYCOPLASMA-HYOPNEUMONIAE
DNA-SEQUENCE DETERMINATION AND BIOCHEMICAL-ANALYSIS OF THE IMMUNOGENIC PROTEIN P36, THE LACTATE-DEHYDROGENASE (LDH) OF MYCOPLASMA-HYOPNEUMONIAE
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DOI：
10.1099/00221287-139-2-317
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发表时间：
1993-02-01
期刊：
影响因子：
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通讯作者：
FREY, J
中科院分区：
文献类型：
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作者：
HALDIMANN, A;NICOLET, J;FREY, J
已经确定了肺炎支原体早期特异性免疫原蛋白P36基因的DNA序列。将P36的DNA序列和推导出的氨基酸序列与数据库中已知的基因和蛋白质进行比较，表明P36是一种l -乳酸脱氢酶（LDH） （EC 1.1.1.27）。对克隆基因在大肠杆菌中表达的蛋白P36进行生化分析，证实P36具有l -乳酸脱氢酶活性。因此肺炎支原体的蛋白P36被称为LDH及其基因LDH。肺炎支原体乳酸脱氢酶被证明含有其他细菌种类乳酸脱氢酶的典型结构域。然而，免疫学上，我们已经证明抗肺炎支原体LDH的多克隆抗体不会与相关LDH交叉反应，并且对肺炎支原体表现出高特异性。ldh基因前面有几个典型的- 10序列，在原核生物的启动子中发现，但缺乏- 35序列。然而，富含A + T的序列位于- 10框之前，这表明与其他细菌物种相比，肺炎支原体中参与转录起始及其调控的因子可能不同，但假定的核糖体结合位点似乎是保守的。
The DNA sequence of the gene encoding the early and specific immunogenic protein P36 of Mycoplasma hyopneumoniae has been determined. Comparison of the DNA sequence and the deduced amino acid sequence of P36 with known genes and proteins in data banks indicated that P36 is a L-lactate dehydrogenase (LDH) (EC 1.1.1.27). Biochemical analysis of protein P36 expressed from the cloned gene in Escherichia coli confirmed that P36 has L-lactate dehydrogenase activity. Protein P36 of M. hyopneumoniae therefore is termed LDH and its gene ldh. M. hyopneumoniae LDH was shown to contain the typical domains of LDH of other bacterial species. Immunologically however, we have shown that polyclonal antibodies against M. hyopneumoniae LDH do not cross-react with related LDH and show high specificity for M. hyopneumoniae. The ldh gene is preceded by several typical - 10 sequences found in promoters of prokaryotes, but lacks the - 35 sequence. Sequences rich in A + T, however, precede the - 10 boxes, suggesting that factors involved in transcription initiation and their regulation may be different in M. hyopneumoniae compared to other bacterial species, but the putative ribosome binding site seems to be conserved.