State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Science, Xujiaping 1, Lanzhou, Gansu 730046, PR China.
Vet Parasitol. 2012 Jul 6;187(3-4):421-30. doi: 10.1016/j.vetpar.2012.01.021. Epub 2012 Jan 21.
A new gene of Babesia sp. BQ1 (Lintan) (BQP35) was cloned by screening a merozoite cDNA expression library with infected sheep serum and using rapid amplification of cDNA ends (RACE). The nucleotide sequence of the cDNA was 1140bp with an open reading frame (ORF) of 936bp encoding a 35-kDa predicted polypeptide with 311 amino acid residues. Comparison of BQP35 cDNA and genomic DNA sequences showed that BQP35 does not possess an intron. Recombinant BQP35 (rBQP35), expressed in a prokaryotic expression system, showed abnormally slow migration on SDS-PAGE. Gel shifting, amino acid sequence and in silico disorder region prediction indicated that BQP35 protein has characteristics of intrinsically unstructured proteins (IUPs). This is the first description of such proteins in the Babesia genus. BQP35 induced antibodies production as early as one week after Babesia sp. BQ1 (Lintan) infection in sheep. No cross-reaction was observed with sera from sheep infected with other ovine piroplasms dominant in China, except with Babesia sp. Tianzhu. The interest of BQP35 as a diagnostic antigen is discussed.
通过用感染绵羊血清筛选裂殖子 cDNA 表达文库,并用快速扩增 cDNA 末端(RACE)技术,克隆了一株新的绵羊巴贝斯虫 BQ1(Lintan)(BQP35)基因。cDNA 的核苷酸序列为 1140bp,含有一个 936bp 的开放阅读框(ORF),编码一个 35kDa 的预测多肽,含有 311 个氨基酸残基。BQP35 cDNA 和基因组 DNA 序列的比较表明,BQP35 不含内含子。在原核表达系统中表达的重组 BQP35(rBQP35)在 SDS-PAGE 上显示出异常缓慢的迁移。凝胶迁移、氨基酸序列和计算无序区预测表明,BQP35 蛋白具有固有无规则蛋白(IUPs)的特征。这是在巴贝斯虫属中首次描述此类蛋白。在绵羊感染巴贝斯虫 BQ1(Lintan)后一周,BQP35 即可诱导抗体产生。与中国流行的其他绵羊血孢子虫感染的血清无交叉反应,除了与天祝巴贝斯虫外。讨论了 BQP35 作为诊断抗原的意义。