随机扩增多态DNA技术在鼠类种属来源判定中的应用研究

Application of random amplified polymorphic DNA for species identification and origin determination of imported rodents

目的研究如何运用随机扩增多态DNA(RAPD)技术判定鼠类种属及来源。方法取鼠尾组织制备DNA基因组,并进行RAPD扩增,以强带差异来区分鼠类遗传多态性。结果建立了褐家鼠DNA快速制备方法,并获得基因图谱。根据120条引物各自表现出的不同多态性,合成一组褐家鼠的通用引物,定名为P1(5′-TCG GCG GTT C-3′)、P2(5′-GAGAGC CGT C-3′)、P3(5′-ATG GCA TCT C-3′)和P4(5′-CTT GGC ACG A-3′),结果显示,该组引物对褐家鼠在目的片段(925 bp)处均有良好的扩增。结论分析个体DNA序列同源程度判断遗传距离大小,可鉴别鼠的来源是否为输入性鼠类,对追溯某种鼠类扩散途径和历史以及预警鼠传疾病提供科学依据。

Objective To use random amplified polymorphic DNA （RAPD） for species identification and origin determination of imported rodents. Methods Genetic DNA of rodents was extracted from tail tissue specimens and amplified by RAPD. Genetic polymorphisms in rodents were distinguished by differences in strong RAPD banding patterns. Results A quick DNA preparation method for sewer rats and their genetic maps were established. In accordance with different polymorphisms of 120 primers, a set of universal primers for sewer rats were synthesized, designated P1 （5＇-TCG GCG GTT C-3＇）, P2 （5＇ -GAG AGC CGT C-3＇ ）, P3 （5＇ -ATG GCA TCT C-3＇ ）, and P4 （5＇ -CTT GGC ACG A-3＇ ）. Amplification of a target fragment （925 bp） in sewer rats using the universal primers all yielded good results. Conclusion Analysis of the degree of homology between individual DNA sequences and determination of their genetic distance allow identifying the origin of rodents as imported or not. The obtained data provide a reference for tracing the dispersal path and history of rodents and detecting early-warning signs of rodent-borne diseases.