EVALUATING THE USE OF THE MULTILOCUS GENETIC MARKER “5S RDNA” FOR THE DISCRIMINATION OF DIFFERENT SPECIES OF FAMILY LETHRINIDAE IN RED SEA, EGYPT

Ghada Mohammed-ElSherif, A. Galal-Khallaf, A. Osman, S. El-Nabi, K. Mohammed-Geba
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Abstract

Family Lethrinidae (Sho´our fishes, Shaari fishes) is one of the key families in the Red Sea and Indian Ocean fisheries. Some of its species were recorded in the International Union for Conservation of Nature (IUCN) Red List of species as endangered. In order to develop simple genetic marker that aid their rapid identification, fin clips of four lethrinid species were collected from local fish markets in Hurghada city. These species were Lethrinus lentjan, L. harak, L. microdon, and L. mahsena. DNA was extracted from these samples and subjected to polymerase chain reaction (PCR) amplification for the mitochondrial 12S ribosomal RNA (rRNA) and the nuclear 5S rRNA genes. The 12S rRNA PCR amplicons were sequenced. GenBank comparison and phylogenetic analysis for the 12S rRNA gene sequences resulted in clear identification for the sampled species. Meanwhile, a simple, 2% agarose gel electrophoresis for the 5S rDNA ran in a low voltage for 2 hours resulted in variable intraspecific patterns. These patterns appeared as 3 PCR bands of approximately 500 base pair (bp) in length for L. lentjan; 2 PCR bands at 500 and 400 bp for L. harak; single PCR band at 500 bp for L. microdon; and 3 bands at about 750, 500, and 400 bp for L. mahsena. Band separation was clear enough. 5S rDNA exhibited an accurate, interspecific, and easy-to-use genetic marker for identification of different lethrinid species covered by the current study.
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利用多位点遗传标记“5s rdna”对埃及红海地区不同种类蠓科昆虫的鉴别评价
Lethrinidae科(shoo ' our鱼类,Shaari鱼类)是红海和印度洋渔业的关键科之一。它的一些物种被列入国际自然保护联盟(IUCN)濒危物种红色名录。为了开发简单的遗传标记,帮助快速鉴定它们,在赫尔格达市当地鱼市收集了四种杀虫剂的鱼鳍片段。这些种分别是Lethrinus lentjan、L. harak、L. microdon和L. mahsena。从这些样本中提取DNA,对线粒体12S核糖体RNA (rRNA)和细胞核5S rRNA基因进行聚合酶链反应(PCR)扩增。对12S rRNA PCR扩增子进行测序。通过GenBank比对和12S rRNA基因序列的系统发育分析,确定了该样本的种类。同时,对5S rDNA进行简单的2%琼脂糖凝胶电泳,在低压下运行2小时,结果显示出不同的种内模式。这些模式为3个PCR条带,长度约为500个碱基对(bp);L. harak在500和400 bp处有2条PCR条带;单PCR条带长度为500 bp;L. mahsena在约750、500和400 bp处有3个波段。波段分离足够清晰。5S rDNA是一种准确、种间和易于使用的遗传标记,可用于本研究涵盖的不同杀虫剂种类的鉴定。
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