盐水微藻杜氏藻中18S rDNA基因的新排列及其ITS区。

Mohammad A Hejazi, Abolfazl Barzegari, Nahid Hosseinzadeh Gharajeh, Mohammad S Hejazi
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引用次数: 44

摘要

比较18S rDNA基因序列是一种很有前途的生物鉴定和分类方法。根据18S rDNA基因的大小、内含子的数量和位置,对不同杜氏藻物种进行了分子鉴定和区分。目前已经报道了三种类型的18S rDNA结构:大小为~1770 bp的基因不含任何内含子,大小为~2170 bp的基因在5'端附近包含一个内含子,大小为~2570 bp的基因在5'和3'端附近包含两个内含子。本文报道了伊朗盐湖杜氏菌(ABRIINW-M1/2)中18S rDNA基因的内含子定位和核苷酸序列。用属特异性引物进行PCR扩增,产生约2170 bp的DNA条带,与盐藻18S rDNA基因相似,该基因在5'端附近仅含1个内含子。同时,该基因的序列组成显示在我们的分离物的5'端附近缺乏任何内含子。此外,由于在3'端附近存在440 bp的DNA片段,观察到另一个变化。因此,该分离物的18S rDNA基因明显不同于D. salina和迄今报道的其他杜氏菌。此外,ITS区域序列分析显示该区域与已有报道的物种相比具有多样性。该分离物的18S rDNA和ITS序列已提交至NCBI数据库,加入号分别为EU678868和EU927373。该菌株的最佳生长速率出现在1 M NaCl的盐度水平。在强光(400 μ mol光子M -2 s-1)、高盐(4 M NaCl)和缺乏硝酸盐和磷酸盐营养的胁迫条件下,15 d后的类胡萝卜素含量最高可达240 ng/ cells。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Introduction of a novel 18S rDNA gene arrangement along with distinct ITS region in the saline water microalga Dunaliella.

Comparison of 18S rDNA gene sequences is a very promising method for identification and classification of living organisms. Molecular identification and discrimination of different Dunaliella species were carried out based on the size of 18S rDNA gene and, number and position of introns in the gene. Three types of 18S rDNA structure have already been reported: the gene with a size of ~1770 bp lacking any intron, with a size of ~2170 bp consisting one intron near 5' terminus, and with a size of ~2570 bp harbouring two introns near 5' and 3' termini. Hereby, we report a new 18S rDNA gene arrangement in terms of intron localization and nucleotide sequence in a Dunaliella isolated from Iranian salt lakes (ABRIINW-M1/2). PCR amplification with genus-specific primers resulted in production of a ~2170 bp DNA band, which is similar to that of D. salina 18S rDNA gene containing only one intron near 5' terminus. Whilst, sequence composition of the gene revealed the lack of any intron near 5' terminus in our isolate. Furthermore, another alteration was observed due to the presence of a 440 bp DNA fragment near 3' terminus. Accordingly, 18S rDNA gene of the isolate is clearly different from those of D. salina and any other Dunaliella species reported so far. Moreover, analysis of ITS region sequence showed the diversity of this region compared to the previously reported species. 18S rDNA and ITS sequences of our isolate were submitted with accesion numbers of EU678868 and EU927373 in NCBI database, respectively. The optimum growth rate of this isolate occured at the salinity level of 1 M NaCl. The maximum carotenoid content under stress condition of intense light (400 mumol photon m-2 s-1), high salinity (4 M NaCl) and deficiency of nitrate and phosphate nutritions reached to 240 ng/cell after 15 days.

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