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利用DNA序列变异研究恶性疟原虫遗传多样性的方法。

Approaches to studying genetic diversity of Plasmodium falciparum using a DNA sequence variation.

作者信息

Robson K J

机构信息

MRC Molecular Haematology Unit, John Radcliffe Hospital, Headington, Oxford, UK.

出版信息

Parassitologia. 1993 Jul;35 Suppl:91-4.

PMID:7901832
Abstract

Sequence variation in the calmodulin and thrombospondin related anonymous protein (TRAP) genes has been examined following amplification using the polymerase chain reaction (PCR). The intron of the calmodulin gene has four repeating motifs which vary in length: three of these contain a dinucleotide repeat, dA-dT, while the fourth is a pentameric repeating unit dA-dT-dA-dT-dT. These DNA polymorphisms can be applied to the study of parasite populations in mixed infections and in strain identification. The known sequence diversity in the TRAP gene has been converted to an RFLP analysis following PCR amplification. Comparisons between the chemical cleavage method and the direct sequencing of PCR amplified products of the TRAP gene, suggest that the latter method is preferable when analysing sequences as variant as TRAP.

摘要

利用聚合酶链反应(PCR)进行扩增后,对钙调蛋白和血小板反应蛋白相关无名蛋白(TRAP)基因中的序列变异进行了检测。钙调蛋白基因的内含子有四个长度各异的重复基序:其中三个含有二核苷酸重复序列dA-dT,而第四个是五聚体重复单元dA-dT-dA-dT-dT。这些DNA多态性可应用于混合感染中寄生虫群体的研究以及菌株鉴定。TRAP基因中已知的序列多样性在PCR扩增后已转化为限制性片段长度多态性(RFLP)分析。对TRAP基因PCR扩增产物的化学切割法和直接测序法进行比较表明,在分析像TRAP这样变异的序列时,后一种方法更可取。

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