CAJ | 학술논문

基于核糖体DNA第一与第二转录间隔序列以及5.8S序列,以分离自广州动物园大熊猫体内的蛔虫为研究对象,用保守引物NC_5和NC_2对核糖体DNA（rDNA）的内转录间隔区ITS-1,ITS-2及5.8S序列进行PCR扩增,扩增后的片段纯化后克隆至pGEM-Teasy载体,重组质粒通过菌液PCR鉴定后,对阳性菌落进行序列测定及分析,鉴定大熊猫蛔虫的种类。结果显示,目的片段总长为910 bp,2个不同样品之间的ITS及5.8S序列没有差异,与GenBank~（TM）中的拜林蛔线虫（Baylisascaris transfuga）、猪蛔虫（Ascaris suum）和人蛔虫（Ascaris lumbricoides）的ITS序列相似性分别为96.6%、82.9%和82.7%。结果表明,此次分离的大熊猫蛔线虫可能为拜林蛔线虫。
기우핵당체DNA제일여제이전록간격서렬이급5.8S서렬,이분리자엄주동물완대웅묘체내적회충위연구대상,용보수인물NC_5화NC_2대핵당체DNA（rDNA）적내전록간격구ITS-1,ITS-2급5.8S서렬진행PCR확증,확증후적편단순화후극륭지pGEM-Teasy재체,중조질립통과균액PCR감정후,대양성균락진행서렬측정급분석,감정대웅묘회충적충류。결과현시,목적편단총장위910 bp,2개불동양품지간적ITS급5.8S서렬몰유차이,여GenBank~（TM）중적배림회선충（Baylisascaris transfuga）、저회충（Ascaris suum）화인회충（Ascaris lumbricoides）적ITS서렬상사성분별위96.6%、82.9%화82.7%。결과표명,차차분리적대웅묘회선충가능위배림회선충。
To identify the species of ascarid samples isolated from a giant panda in Guangzhou Zoo,China,we designed a pair of conserved primers（NC2/NC5） to amplify the first and second internal transcribed spacers（ITS） and 5.8S of ribosomal DNA（5.8S rDNA）.The ITS rDNA was amplified by PCR from the ascarid samples.The amplicons were then purified and cloned into pGEM - T Easy vector.After being identified by PCR amplification and restriction digestion,the positive clones were sequenced and the sequences were compared with those of Baylisascaris transfuga,Ascaris sum and Ascaris lumbricoides available in GenBank~（TM）.The results revealed that the two ascarid specimens had identical ITS and 5.8S rDNA sequences of 910bp length.The ITS sequences of the ascarid nematodes from the giant panda were significantly different from those of Baylisascaris transfuga,Ascaris sum,and Ascaris lumbricoides,with similarities of 96.6%,82.9%,and 82.7%,respectively. The ascarid nematodes from the giant panda differed from the species listed above and their ITS sequence had not been previously reported.We provisionally identify the species as Baylisascaris transfuga,the species showing greatest similarity.