Milhausen M, Nelson R G, Parsons M, Newport G, Stuart K, Agabian N
Mol Biochem Parasitol. 1983 Nov;9(3):241-54. doi: 10.1016/0166-6851(83)90100-7.
Variant surface glycoproteins (VSGs) were isolated from variant antigen types (VATs) of the IsTat 1 serodeme. Molecular weight and isoelectric focusing analysis demonstrate that seven early VSGs possess properties generally attributed to VSGs isolated from other trypanosome serodemes. Six of the seven VSGs characterized are distinct from one another, while two (D and 1) appear identical. The presence of VSG specific mRNA in corresponding VATs was demonstrated by in vitro translation of RNA from each of the VATs, followed by immunoprecipitation with homologous and heterologous antisera. Hybridization of VSG cDNA clones with RNA from each VAT confirm that VSG mRNA is present only in homologous VATs and verifies the transcriptional control of these VSG genes. The two VATs D and 1 express indistinguishable VSGs by a variety of biochemical criteria, as well as by reactivity with 24 monoclonal antibodies. The VSG mRNAs in VATs D and 1 also appear identical. However, this identity is not reflected at the genomic level. Data is presented which establishes that DNA rearrangements can occur around both expressed and non-expressed VSG genes without qualitatively affecting VSG gene expression.
从IsTat 1血清型的变异抗原类型(VATs)中分离出变异表面糖蛋白(VSGs)。分子量和等电聚焦分析表明,七种早期VSGs具有通常归因于从其他锥虫血清型中分离出的VSGs的特性。所鉴定的七种VSGs中有六种彼此不同,而两种(D和1)似乎相同。通过对每个VAT的RNA进行体外翻译,然后用同源和异源抗血清进行免疫沉淀,证明了相应VAT中存在VSG特异性mRNA。VSG cDNA克隆与每个VAT的RNA杂交证实,VSG mRNA仅存在于同源VAT中,并验证了这些VSG基因的转录控制。根据多种生化标准以及与24种单克隆抗体的反应性,VAT D和1表达难以区分的VSGs。VAT D和1中的VSG mRNA也似乎相同。然而,这种同一性在基因组水平上并未体现出来。所提供的数据表明,DNA重排可发生在已表达和未表达的VSG基因周围,而不会在质量上影响VSG基因的表达。