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首页> 外文期刊>Infection and immunity >Decorin-Binding Protein of Borrelia burgdorferi Is Encoded within a Two-Gene Operon and Is Protective in the Murine Model of Lyme Borreliosis
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Decorin-Binding Protein of Borrelia burgdorferi Is Encoded within a Two-Gene Operon and Is Protective in the Murine Model of Lyme Borreliosis

机译：伯氏疏螺旋体的Decorin结合蛋白被编码在两个基因的操纵子中，并在莱姆疏螺旋体的鼠模型中具有保护作用。

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Isolated outer membranes of Borrelia burgdorferi were used in immunoblotting experiments with sera from immune mice to identify new putative Lyme disease vaccine candidates. One immunoreactive polypeptide migrated on polyacrylamide gels just proximal to outer surface protein C and comigrated with [3H]palmitate-labeled polypeptides. A degenerate oligonucleotide primer based upon internal amino acid sequence information was used to detect the corresponding gene within a B. burgdorferi total genomic library. The relevant open reading frame (ORF) encoded a polypeptide comprised of a 24-amino-acid putative signal peptide terminated by LLISC, a probable consensus sequence for lipoprotein modification, and a mature protein of 163 amino acids. Immunoblots of a recombinant fusion protein corresponding to this ORF supported the idea that the encoded protein was a previously reported decorin-binding protein (DBP) of B. burgdorferi N40 (B. P. Guo, S. J. Norris, L. C. Rosenberg, and M. H??k, Infect. Immun. 63:3467–3472, 1995). However, further DNA sequencing revealed the presence of a second ORF, designated ORF-1, whose termination codon was 119 bp upstream of the dbpgene. ORF-1 also encoded a putative lipoprotein with a mature length of 167 amino acids. Northern blots, Southern blots, and primer extension analyses indicated that ORF-1 and dbp comprised a two-gene operon located on the 49-kb linear plasmid. Both proteins, which were 40% identical and 56% similar, partitioned into Triton X-114 detergent extracts of B. burgdorferi isolated outer membranes. Mice infected with B. burgdorferi produced high titers of antibodies against the ORF-1-encoded protein and DBP during both early and later stages of chronic infection. Both DBP and the ORF-1-encoded protein were sensitive to proteinase K treatment of intact borreliae, suggesting that they were surface exposed. In active immunization experiments, 78% of mice immunized with recombinant DBP were immune to challenge. While it is not clear whether the two lipoproteins encoded by the ORF-1-dbp operon have analogous decorin-binding functions in vivo, the combined studies implicate DBP as a new candidate for a human Lyme disease vaccine.
机译：分离的 Borrelia burgdorferi 外膜用于免疫小鼠血清的免疫印迹实验，以鉴定新的假定的莱姆病候选疫苗。一种免疫反应性多肽迁移到聚丙烯酰胺凝胶上，紧靠外表面蛋白C，并与[ 3 H]棕榈酸酯标记的多肽迁移。基于内部氨基酸序列信息的简并寡核苷酸引物用于检测 B内的相应基因。 burgdorferi 全基因组文库。相关的开放阅读框（ORF）编码的多肽包含由LLISC终止的24个氨基酸推定的信号肽，可能的脂蛋白修饰共有序列以及163个氨基酸的成熟蛋白。对应于该ORF的重组融合蛋白的免疫印迹支持这样的想法，即编码的蛋白是先前报道的 B的核心蛋白聚糖结合蛋白（DBP）。 burgdorferi N40（B。P. Guo，S。J. Norris，L。C. Rosenberg和M. H？k，Infect。Immun。63：3467–3472，1995）。但是，进一步的DNA测序表明存在第二个ORF，称为ORF-1，其终止密码子在 dbp 基因上游119 bp。 ORF-1还编码一个假定的脂蛋白，成熟长度为167个氨基酸。 Northern印迹，Southern印迹和引物延伸分析表明，ORF-1和 dbp 包含一个位于49kb线性质粒上的双基因操纵子。两种蛋白质（分别具有40％的相似性和56％的相似性）分配到Triem X-114 B洗涤剂提取物中。 burgdorferi 分离的外膜。感染了 B的小鼠。 burgdorferi 在慢性感染的早期和晚期均产生高滴度的针对ORF-1编码的蛋白质和DBP的抗体。 DBP和ORF-1编码的蛋白均对完整疏螺旋体的蛋白酶K处理敏感，表明它们是表面暴露的。在主动免疫实验中，重组DBP免疫的小鼠中有78％对攻击免疫。虽然尚不清楚由ORF-1- dbp 操纵子编码的两种脂蛋白在体内是否具有类似的除芯蛋白结合功能，但联合研究表明DBP作为人莱姆病疫苗的新候选者。

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