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首页> 外文期刊>Aquaculture Research >Characterization of growth-related genes in the south-western Atlantic pink shrimp Farfantepenaeus paulensis (Perez-Farfante 1967) through a modified DDRT-PCR protocol.
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Characterization of growth-related genes in the south-western Atlantic pink shrimp Farfantepenaeus paulensis (Perez-Farfante 1967) through a modified DDRT-PCR protocol.

机译:通过改进的DDRT-PCR方案表征西南大西洋粉红虾Farfantepenaeus paulensis(Perez-Farfante 1967)中与生长相关的基因。

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摘要

An exploratory methodology has been used to identify genes related to growth of the shrimp Farfantepenaeus paulensis (Perez-Farfante, 1967). Six hundred postlarvae, with an average (+or-SD) initial weight of 0.022 g (+or-0.008), were reared for 60 days in salinity 30 g L-1. In the end, the 15 heaviest (>1.2 g) and the 15 lightest (<0.6 g) animals were frozen in liquid nitrogen. A modified differential display reverse transcription-polymerase chain reaction technique was used to generate expression profiles for all individuals. The resulting cDNA from reverse transcription were amplified by pairing 31 arbitrary primers with a reverse primer anchored to the cDNA tails. From the comparison between the expression profiles of the organisms in the two size classes, differences could be pinpointed. Bands of interest were collected, purified, cloned and sequenced. Despite the relatively small number of primers used, the methodology allowed the identification of three genes, not described previously for the investigated species and possibly related to growth. These partial sequences are likely to belong to genes that code for myosin heavy chain, cyclophilin and haemocyanin as revealed through an amino acid similarity search conducted using the blastp on-line tool.
机译:已经使用一种探索性方法来鉴定与虾Farfantepenaeus paulensis生长有关的基因(Perez-Farfante,1967)。以盐度为30 g L-1饲养60天平均初始重量(+或-SD)为0.022 g(+-0.008)的幼虫。最后,将15只最重(> 1.2 g)和15只最轻(<0.6 g)的动物冷冻在液氮中。改良的差异显示逆转录聚合酶链反应技术用于生成所有个体的表达谱。通过将31个任意引物与锚定在cDNA尾部的反向引物配对来扩增从逆转录得到的cDNA。通过比较两种大小类别的生物的表达谱,可以查明差异。收集感兴趣的条带,进行纯化,克隆和测序。尽管使用的引物数量相对较少,但该方法仍可鉴定三个基因,这些基因先前未针对所研究物种进行过描述,可能与生长有关。这些部分序列可能属于编码肌球蛋白重链,亲环蛋白和血蓝蛋白的基因,这是通过使用blastp在线工具进行的氨基酸相似性搜索揭示的。

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