Cloning and DNA sequencing of the dextranase inhibitor gene (dei) from Streptococcus sobrinus.
Cloning and DNA sequencing of the dextranase inhibitor gene (dei) from Streptococcus sobrinus.
复制标题
远缘链球菌葡聚糖酶抑制剂基因 (dei) 的克隆和 DNA 测序。
DOI：
10.1128/jb.176.23.7213-7222.1994
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发表时间：
1994
影响因子：
3.2
通讯作者：
Curtiss3rd,R
中科院分区：
文献类型：
--
作者：
Sun,JW;Wanda,SY;Camilli,A;Curtiss3rd,R
远缘链球菌UAB 66（血清型g）的一些葡聚糖酶缺陷（Dex-）突变体合成抑制葡聚糖酶活性的物质（S. Y. Wanda，A. Camilli，H. M. Murchison和R. Curtiss III，J. Bacteriol. 176：7206-7212，1994）。由Dex突变体UAB 108产生的这种物质被指定为葡聚糖酶抑制剂（Dei），并被鉴定为蛋白质。将来自UAB 108的Dei基因（dei）克隆到pACYC 184中以产生pYA 2651，然后将其用于产生几个亚克隆（pYA 2653至pYA 2657）。通过使用pYA 2653的Tn 5seq 1转座子诱变来确定dei的DNA序列。dei的开放阅读框长990 bp。它编码38个氨基酸的信号肽和292个氨基酸的成熟Dei蛋白，分子量为31，372。Dei的氨基酸序列与葡糖基转移酶和葡聚糖结合蛋白具有不同程度的相似性，并含有可能参与葡聚糖结合的A和C重复单元。Southern杂交结果表明，UAB 108的dei探针与S. sobrinus（血清型d和g）DNA，在S. downei（血清型h）和S.而对其他变形链球菌群的DNA完全不敏感。
Some dextranase-deficient (Dex-) mutants of Streptococcus sobrinus UAB66 (serotype g) synthesize a substance which inhibits dextranase activity (S.-Y. Wanda, A. Camilli, H. M. Murchison, and R. Curtiss III, J. Bacteriol. 176:7206-7212, 1994). This substance produced by the Dex- mutant UAB108 was designated dextranase inhibitor (Dei) and identified as a protein. The Dei gene (dei) from UAB108 has been cloned into pACYC184 to yield pYA2651, which was then used to generate several subclones (pYA2653 to pYA2657). The DNA sequence of dei was determined by using Tn5seq1 transposon mutagenesis of pYA2653. The open reading frame of dei is 990 bp long. It encodes a signal peptide of 38 amino acids and a mature Dei protein of 292 amino acids with a molecular weight of 31,372. The deduced amino acid sequence of Dei shows various degrees of similarity with glucosyltransferases and glucan-binding protein and contains A and C repeating units probably involved in glucan binding. Southern hybridization results showed that the dei probe from UAB108 hybridized to the same-size fragment in S. sobrinus (serotype d and g) DNA, to a different-size fragment in S. downei (serotype h) and S. cricetus (serotype a), and not at all to DNAs from other mutans group of streptococci.