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首页> 外文期刊>Infection and immunity >Identification, Characterization, and Expression of Three New Members of the Borrelia burgdorferi Mlp (2.9) Lipoprotein Gene Family
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Identification, Characterization, and Expression of Three New Members of the Borrelia burgdorferi Mlp (2.9) Lipoprotein Gene Family

机译：伯氏疏螺旋体Mlp（2.9）脂蛋白基因家族的三个新成员的鉴定，表征和表达

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We previously reported on the existence of a family of lipoprotein genes, designated 2.9 lipoprotein genes, encoded in at least seven versions on the circular (supercoiled) cp32 and cp18 plasmids ofBorrelia burgdorferi 297. A distinguishing feature of the 2.9 lipoproteins were highly similar signal sequences but variable mature polypeptides that segregated into two antigenic classes. Further screenings of B. burgdorferi 297 genomic libraries led to the identification of three additional 2.9 lipoprotein genes, renamed herein mlp, for multicopy lipoprotein genes. Computer analyses and immunoblotting revealed that Mlp-9 segregated with the antigenic class I lipoproteins, whereas Mlp-8 and Mlp-10 were members of class II. Northern blotting showed that all three of themlp genes were expressed when B. burgdorferiwas cultivated in vitro at 34°C, although mlp-9 andmlp-10 transcripts were expressed at very low levels. Additional combined immunoblotting and comparative reverse transcription-PCR analyses performed on borreliae cultivated in vitro at 23, 34, or 37°C indicated that although Mlp-8 was substantially more abundant than Mlp-9 or Mlp-10, all three of the mlpgenes were upregulated during B. burgdorferi replication at 37°C. Expression of the same three lipoproteins was further enhanced upon growth of the spirochetes within dialysis membrane chambers (DMCs) implanted intraperitoneally in rats (i.e., spirochetes in a mammalian host-adapted state), suggesting that temperature alone did not account for maximal upregulation of the mlp genes. That certainmlp genes are likely expressed during the growth ofB. burgdorferi in mammalian tissues was supported by findings of antibodies against all three Mlp lipoproteins in mice after challenge with Ixodes scapularis nymphs harboring B. burgdorferi 297. The combined data suggest that as opposed to being differentially expressed in any reciprocal fashion (e.g., OspA/OspC), at least three mlp genes are simultaneously upregulated by temperature (37°C) and some other mammalian host factor(s). The findings have importance not only for understanding alternative modes of differential antigen expression by B. burgdorferi but also for assessing whether one or more of the Mlp lipoproteins represent new candidate vaccinogens for Lyme disease.
机译：我们之前曾报道过存在一个脂蛋白基因家族，称为2.9脂蛋白基因，至少在 Borrelia burgdorferi 297的圆形（超螺旋）cp32和cp18质粒上编码有至少七个版本。 2.9脂蛋白是高度相似的信号序列，但是可变的成熟多肽被分为两个抗原类别。 B的进一步筛选。 burgdorferi 297个基因组文库导致鉴定出了三个额外的2.9脂蛋白基因，在这里更名为 mlp ，用于多拷贝脂蛋白基因。计算机分析和免疫印迹表明，Mlp-9与抗原性I类脂蛋白分离，而Mlp-8和Mlp-10是II类成员。 Northern印迹显示，当 B时，所有三个 mlp 基因均表达。 burgdorferi 是在34°C体外培养的，尽管 mlp-9 和 mlp-10 转录本的表达水平很低。对在23、34或37°C下体外培养的疏螺旋体进行的其他结合的免疫印迹和比较性逆转录PCR分析表明，尽管Mlp-8比Mlp-9或Mlp-10丰富得多，但所有三个在 B期间，> mlp 基因被上调。 burgdorferi 在37°C下复制。在大鼠腹膜内植入的透析膜腔（DMC）中，螺旋体的生长会进一步增强相同三种脂蛋白的表达（即，处于哺乳动物宿主适应状态的螺旋体），这表明温度本身并不能说明最大的上调。 mlp 基因。某些 mlp 基因可能在 B的生长过程中表达。在带有 B的肩em小突触虫攻击后，针对小鼠中所有三种Mlp脂蛋白的抗体的发现支持了哺乳动物组织中的burgdorferi 。 burgdorferi 297.组合数据表明，与以任何倒数方式（例如OspA / OspC）差异表达相反，至少三个 mlp 基因同时被温度上调（37° C）和其他一些哺乳动物宿主因子。该发现不仅对理解 B的差异抗原表达的替代模式具有重要意义。 burgdorferi ，还用于评估一种或多种Mlp脂蛋白是否代表莱姆病的新候选疫苗致癌物。

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