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7H8/6,一种用于区分恶性疟原虫分离株的多拷贝DNA探针。

7H8/6, a multicopy DNA probe for distinguishing isolates of Plasmodium falciparum.

作者信息

Limpaiboon T, Shirley M W, Kemp D J, Saul A

机构信息

Queensland Institute of Medical Research, Herston, Australia.

出版信息

Mol Biochem Parasitol. 1991 Aug;47(2):197-206. doi: 10.1016/0166-6851(91)90179-a.

Abstract

A method using DNA fingerprinting to distinguish Plasmodium falciparum isolates has been developed using a clone, 7H8/6, isolated by screening a genomic expression library of the Malayan Camp strain with a monoclonal antibody. Unlike other P. falciparum DNA probes that contain internally repetitive sequences, this probe contains a single full-length open reading frame and lacks any repetitive sequences. Chromosome mapping located the hybridizing sequences to most chromosomes. 7H8/6 sequences are located at sites near the subtelomeric regions, although they are further from the telomeres than the subtelomeric repetitive sequences rep20. The probe gave multiple polymorphic bands on Southern blots of genomic DNA allowing all unrelated isolates tested to be readily distinguished. Hybridization patterns of parent and progeny from the genetic cross of two P. falciparum clones showed that most bands were inherited independently and no new bands were generated during the cross. This probe is useful for distinguishing isolates of P. falciparum and also for analyzing relationships between closely related isolates.

摘要

一种利用DNA指纹识别技术区分恶性疟原虫分离株的方法已被开发出来,该方法使用了一个名为7H8/6的克隆,它是通过用单克隆抗体筛选马来亚营地株的基因组表达文库分离得到的。与其他含有内部重复序列的恶性疟原虫DNA探针不同,该探针包含一个单一的全长开放阅读框,并且缺乏任何重复序列。染色体图谱分析将杂交序列定位到大多数染色体上。7H8/6序列位于亚端粒区域附近的位点,尽管它们比亚端粒重复序列rep20离端粒更远。该探针在基因组DNA的Southern印迹上产生多个多态性条带,使得所有测试的无关分离株都能很容易地被区分开来。两个恶性疟原虫克隆的遗传杂交中亲本和子代的杂交模式表明,大多数条带是独立遗传的,并且在杂交过程中没有产生新的条带。该探针可用于区分恶性疟原虫的分离株,也可用于分析密切相关分离株之间的关系。

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