HFJ和MIJ近交系大鼠RAPD特征与比较研究

The Study of Randomly Amplified Polymorphic DNA Characteristic in HFJ and MIJ Inbred Rats

目的利用人工合成的PCR随机引物,对本单位培育的HFJ和MIJ近交系大鼠基因组DNA进行扩增,建立HFJ和MIJ大鼠RAPD扩增特征图谱;选择常用近交系大鼠Lewis和F344作为对照品系,观察HFJ和MIJ大鼠与对照品系大鼠的RAPD多态性。方法用酚-氯仿法分别提取4个不同品系大鼠的基因组DNA,从60条随机引物中筛选能够得到清晰扩增条带的30条引物,对HFJ和MIJ大鼠DNA进行扩增。再从30条引物中随机选择12条,对4个品系大鼠基因组DNA扩增,并对扩增条带多态性比较分析。结果 30条引物对HFJ和MIJ大鼠DNA进行扩增,均能获得1～7个较清晰条带,除引物O～08外,条带的分子大小均在200～1 100 bp之间,两品系的引物O-08扩增结果,均在2 000 bp以上的近原点处扩增出1条清晰的条带。HFJ和MIJ大鼠品系内个体间和两个品系之间,扩增的结果均完全一致;用12条随机引物对4个品系大鼠同时扩增,HFJ和MIJ大鼠扩增结果一致,与Lewis和F344比较,有5个引物出现差异条带。结论用30条随机引物建立了HFJ和MIJ大鼠RAPD扩增特征性图谱。由于两品系来源于同一对Wistar封闭群大鼠,虽然遗传特性有显著差异,但是在所选30条引物扩增时,未发现多态性。两品系与Lewis和F344之间存在多态性条带。

Objective To establish the fingerprint of HFJ and MIJ inbred rats by randomly amplified polymorphic DNA（RAPD） analysis.And to compare the DNA polymorphism of the four inbreeds rats including HFJ,MIJ,Lewis and F344.Methods To isolate rats genomic DNA by standard procedure of phenol/chloroform.Sixty primers were used to amplify rats DNA with RAPD-PCR.Thirty primers could produce clear fragments and were selected to analyze the DNA polymorphism between HFJ and MIJ inbred rats.Twelve primers were selected to analyze the DNA polymorphism among HFJ,MIJ,Lewis and F344 inbred rats.Results 1~7 clear bands were detected by electrophoresis on agarose gel in HFJ and MIJ inbred rats with thirty primers respectively.All amplified bands by random oligonucleotide primers were located between 200 bp^1 100 bp except primer O^08,which located over 2 000 bp and there was a distinct band near the origin of electrophoresis.The results of RAPD indicated that there were identical completely between HFJ and MIJ inbred rats or within the two strains.DNA polymorphisms among HFJ,MIJ,Lewis and F344 inbred rats were studied by RAPD using twelve primers.The results indicated that 5 primers could to obtain difference fragments in all twelve primers.Conclusion With the thirty random primers to establish RAPD fingerprint of HFJ and MIJ inbred rats.Since the two inbred strains originated from a pair of Wistar rats,DNA polymorphism between HFJ and MIJ inbred rats were no detected,although the two inbred strains had remarkable genetic difference.By RAPD analysis,DNA polymorphism among HFJ,MIJ,Lewis and F344 inbred rats were detected.