PCR-based approach for sequencing mitochondrial genomes of decapod crustaceans, with a practical example from kuruma prawn (Marsupenaeus japonicus)
PCR-based approach for sequencing mitochondrial genomes of decapod crustaceans, with a practical example from kuruma prawn (Marsupenaeus japonicus)
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DOI：
10.1007/s10126-003-0036-2
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发表时间：
2004-09-01
影响因子：
3
通讯作者：
Nishida, M
中科院分区：
文献类型：
--
作者：
Yamauchi, MM;Miya, MU;Nishida, M
描述了一种使用 79 个新设计的和 7 个已发表的聚合酶链反应 (PCR) 引物对十足目甲壳类动物的整个线粒体基因组（线粒体基因组）进行测序的方法。该方法包括以下步骤：（1）在2或3个长PCR中扩增整个有丝分裂基因组； (2)以稀释的长PCR产物为模板，以不同的组合使用86个引物，扩增整个有丝分裂基因组中连续、重叠的短片段； (3)利用短PCR产物进行直接循环测序。与使用克隆线粒体 DNA 和引物步移策略的传统方法相比，该策略可以更快速地确定十足目线粒体基因组序列。作为一个实际例子，使用基于 PCR 的方法确定了库鲁玛虾 Marsupenaeus japonicus（甲壳纲：十足目）的线粒体基因组序列。
An approach for sequencing the entire mitochondrial genomes (mitogenomes) of decapod crustaceans using 79 newly designed and 7 published polymerase chain reaction (PCR) primers is described. The approach comprises the following steps: (1) the entire mitogenome is amplified in 2 or 3 long PCRs; (2) the 86 primers are used in different combinations to amplify contiguous, overlapping short segments of the entire mitogenome with the diluted long PCR products as templates; (3) direct cycle sequencing is conducted using the short PCR products. This strategy allows a more rapid determination of decapod mitogenomic sequences than a traditional method using cloned mitochondrial DNA and primer walking strategy. As a practical example, the mitogenomic sequence for a kuruma prawn Marsupenaeus japonicus (Crustacea: Decapoda), was determined using the PCR-based approach.