Takle G B, Young A, Snary D, Hudson L, Nicholls S C
Department of Immunology, St. George's Hospital Medical School, London, U.K.
Mol Biochem Parasitol. 1989 Nov;37(1):57-64. doi: 10.1016/0166-6851(89)90102-3.
An 85-kDa trypomastigote-specific surface antigen gene from Trypanosoma cruzi has been identified by screening a genomic library in lambda gt10 with trypomastigote and epimastigote cDNA. The 1.3-kb genomic clone (pTt34) hybridizes to a single trypomastigote mRNA of 3.7 kb and to multiple bands in genomic Southern blots. Dot-blot experiments show that there are 5-10 copies of this sequence per haploid genome, and these are arranged in a non-tandem manner. pTt34 has been expressed as an anthranilate synthetase fusion protein in Escherichia coli, and inclusion bodies have been used to raise antiserum in rabbits. This antiserum immunoprecipitates a cell surface trypomastigote-specific protein of 85 kDa. The DNA and predicted amino acid sequences of pTt34 are given. Four further clones obtained from a PvuII/HpaI partial genomic library in pUC13 have extended the sequence of the 3' end of pTt34; each of these clones has regions of sequence divergence and each could represent a different member of the gene family.
通过用锥鞭毛体和上鞭毛体cDNA筛选λgt10基因组文库，已鉴定出克氏锥虫一个85 kDa的锥鞭毛体特异性表面抗原基因。1.3 kb的基因组克隆（pTt34）与一条3.7 kb的单一锥鞭毛体mRNA杂交，并与基因组Southern印迹中的多条带杂交。斑点印迹实验表明，每个单倍体基因组中有5 - 10个该序列的拷贝，且这些拷贝以非串联方式排列。pTt34已在大肠杆菌中表达为邻氨基苯甲酸合成酶融合蛋白，其包涵体已用于在兔中制备抗血清。该抗血清免疫沉淀出一种85 kDa的细胞表面锥鞭毛体特异性蛋白。给出了pTt34的DNA和预测的氨基酸序列。从pUC13中的PvuII/HpaI部分基因组文库获得的另外四个克隆扩展了pTt34 3'端的序列；这些克隆中的每一个都有序列差异区域，且每一个都可能代表该基因家族的不同成员。
