克氏锥虫核糖体RNA基因的分子克隆及部分特性分析

Molecular cloning and partial characterization of ribosomal RNA genes from Trypanosoma cruzi.

作者信息

Hernández R, Díaz-de Léon F, Castañeda M

机构信息

Department of Developmental Biology, Universidad Nacional Autónoma de México, Mexico City.

出版信息

Mol Biochem Parasitol. 1988 Jan 15;27(2-3):275-9. doi: 10.1016/0166-6851(88)90047-3.

DOI:10.1016/0166-6851(88)90047-3

PMID:3278230

Abstract

To further analyze the organization of the nuclear rDNA locus in Trypanosoma cruzi, genomic recombinant plasmid clones were constructed and isolated after hybridization with rRNA molecules as hybridization probes. Approximately 11 kilobase pairs from the cistron were cloned in three recombinant plasmids carrying adjacent genomic fragments. Restriction mapping and Southern hybridization experiments performed on these clones indicate the following relative arrangement of the mature rRNA coding sequences: 18S (2.46 kb), S3 (197 b), 24S alpha (2.02 kb), S1 (261 b), 24 beta (1.66 kb), S2 (217 b) and S6 (90 b). Neither S4 (141 b) nor S5 (110 b) sequences were found within these genomic clones. Nevertheless genomic Southerns suggest a linkage of S4 towards the 3' end of this genetic system.

摘要

为了进一步分析克氏锥虫核仁rDNA基因座的组织情况，以rRNA分子作为杂交探针，构建并分离了基因组重组质粒克隆。从顺反子中约11千碱基对被克隆到携带相邻基因组片段的三个重组质粒中。对这些克隆进行的限制性图谱分析和Southern杂交实验表明，成熟rRNA编码序列的相对排列如下：18S（2.46 kb）、S3（197 bp）、24Sα（2.02 kb）、S1（261 bp）、24β（1.66 kb）、S2（217 bp）和S6（90 bp）。在这些基因组克隆中未发现S4（141 bp）和S5（110 bp）序列。然而，基因组Southern杂交表明S4与该遗传系统的3'端相连。