Reverse transcription of trypanosome variable antigen mRNAs initiated by a specific oligonucleotide primer.
Reverse transcription of trypanosome variable antigen mRNAs initiated by a specific oligonucleotide primer.
复制标题
由特定寡核苷酸引物启动锥虫可变抗原 mRNA 的逆转录。
DOI：
10.1073/pnas.80.6.1536
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发表时间：
1983
影响因子：
11.1
通讯作者：
Richards,FF
中科院分区：
文献类型：
--
作者：
Merritt,SC;Tschudi,C;Konigsberg,WH;Richards,FF
非洲锥虫通过连续表达一组相关但高度多样化的蛋白质的不同成员，即变体表面糖蛋白（VSG）来改变其抗原性。我们描述了一种合成的寡核苷酸，它可以特异性地引发总锥虫poly（A）+ RNA中VSG mRNA的逆转录。通过转录产物的cDNA序列分析和单个cDNA与细胞RNA的变体特异性杂交的证明来验证这种引发的特异性。该寡核苷酸引物还用作锥虫基因组DNA文库中这些VSG mRNA上发现的保守序列的探针。在冈比亚锥虫基因组文库中检测到大量引物阳性克隆，而在刚果锥虫基因组DNA文库中检测到很少的阳性信号。
African trypanosomes change their antigenicity by successively expressing different members of a group of related but highly diverse proteins, the variant surface glycoproteins (VSGs). We describe a synthetic oligonucleotide that can prime specifically reverse transcription of VSG mRNA out of total trypanosome poly(A)+ RNA. The specificity of this priming was verified by cDNA sequence analysis of the transcription products and by the demonstration of variant-specific hybridization of the individual cDNAs to cellular RNA. The oligonucleotide primer also was used as a probe for the conserved sequence found on these VSG mRNAs in trypanosome genomic DNA libraries. A large number of primer-positive clones were detected in a Trypanosoma gambiense genomic library, but very few positive signals were found in a library of Trypanosoma congolense genomic DNA.