海洋科学
海洋科學
해양과학
MARINE SCIENCES
2013年
9期
28-36
,共9页
李善策%李勇勇%夏金兰%秦松
李善策%李勇勇%夏金蘭%秦鬆
리선책%리용용%하금란%진송
钝顶节旋藻(Arthrospira platensis)%全基因组提取%PCR%16S rRNA-ITS
鈍頂節鏇藻(Arthrospira platensis)%全基因組提取%PCR%16S rRNA-ITS
둔정절선조(Arthrospira platensis)%전기인조제취%PCR%16S rRNA-ITS
Arthrospira platensis%whole genome DNA extraction%PCR%16S rRNA-ITS
本研究从实验室保藏的节旋藻(Arthrospira)藻种出发,挑取形态不同的单藻丝体进行纯化培养,采用6种方法进行全基因组DNA提取的比较研究,而后以16S rRNA-ITS区基因作为分子标记对藻株进行相关序列测定和分子系统进化分析。结果表明,冻融 CTAB 法能够提取出包含染色体外 DNA 在内的节旋藻全基因组,高质量样品可以满足分子生物学实验要求;分子系统研究表明,纯化藻株皆为钝顶节旋藻,节旋藻与螺旋藻在分子鉴定中属间差异明显。
本研究從實驗室保藏的節鏇藻(Arthrospira)藻種齣髮,挑取形態不同的單藻絲體進行純化培養,採用6種方法進行全基因組DNA提取的比較研究,而後以16S rRNA-ITS區基因作為分子標記對藻株進行相關序列測定和分子繫統進化分析。結果錶明,凍融 CTAB 法能夠提取齣包含染色體外 DNA 在內的節鏇藻全基因組,高質量樣品可以滿足分子生物學實驗要求;分子繫統研究錶明,純化藻株皆為鈍頂節鏇藻,節鏇藻與螺鏇藻在分子鑒定中屬間差異明顯。
본연구종실험실보장적절선조(Arthrospira)조충출발,도취형태불동적단조사체진행순화배양,채용6충방법진행전기인조DNA제취적비교연구,이후이16S rRNA-ITS구기인작위분자표기대조주진행상관서렬측정화분자계통진화분석。결과표명,동융 CTAB 법능구제취출포함염색체외 DNA 재내적절선조전기인조,고질량양품가이만족분자생물학실험요구;분자계통연구표명,순화조주개위둔정절선조,절선조여라선조재분자감정중속간차이명현。
Single filaments of Arthrospira/Spirulina strains were picked up and axenicly cultured. Six protocols for ex-traction of whole genome DNA of the Arthrospira strains were comparatively studied. The phylogenetic analysis of 16S rRNA-ITS (internally transcribed spacer) gene sequences as molecular markers were conducted. The results show that the method of Freezing-thawing CTAB was effective in extracting extra-genomic DNA, not chromasomal from the tested strains. The extracted DNA containing excellent overall quality and high molecular weight can be directly used for molecular biology experiments. The molecular phylogenetic dendrogram indicates that the strains used in this research were all Arthrospira platensis, which were significantly different from strains of Spirulina in molecu-lar classification and identification in cyanobacterial genus.