Oligonucleotide primers for targeted amplification of single-copy nuclear genes in apocritan Hymenoptera.
Oligonucleotide primers for targeted amplification of single-copy nuclear genes in apocritan Hymenoptera.
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DOI：
10.1371/journal.pone.0039826
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Niehuis O
中科院分区：
文献类型：
--
作者：
Hartig G;Peters RS;Borner J;Etzbauer C;Misof B;Niehuis O
来自大型多样性昆虫序膜翅目（锯蝇，蜜蜂，黄蜂和蚂蚁）的核苷酸序列数据在分类学上分散，仍然不足以重建良好的系统发育树。通过靶向PCR获得的综合多个基因数据集的分析可以为此问题提供成本效益的解决方案。然而，寡核苷酸引物可用于在多种膜翅目种类中核基因的PCR扩增。
在这里，我们提出了一套退化的寡核苷酸底漆对，用于从膜翅目中的154个单拷贝核蛋白质编码基因进行PCR扩增。这些引物是从九个膜翅目（七种蚂蚁，蜜蜂和寄生虫鼻虫肾上腺素）的基因组序列数据推断出来的。我们经验测试了这些引物对的随机选择子集，用于从六个膜翅目扩增靶基因，代表Chrysididae，Crabronidae，Gasteruptiidae，Leucostiidae，Leucostiae，Pompilidae和Stephanidae家族。根据我们的结果，我们估计这些引物适用于在广泛的apocritan膜翅目（即所有具有黄蜂的膜翅目）和尖齿膜翅目（即，尤其是具有apocritan temps apocritan Wasps）的大量核基因（即，所有具有黄蜂的膜翅目）的核基因（即刺痛者）。
放大的核苷酸序列是（a）单拷贝基因的概率很高，（b）在低财务成本下很容易产生，尤其是与系统基因组方法相比，（c）通过另外提供的测序引物，轻松测序（d）适合解决广泛的系统发育问题并通过条形码来帮助快速物种鉴定，因为许多扩增子都包含外显子和快速发展的内含子核苷酸。
Published nucleotide sequence data from the mega-diverse insect order Hymenoptera (sawflies, bees, wasps, and ants) are taxonomically scattered and still inadequate for reconstructing a well-supported phylogenetic tree for the order. The analysis of comprehensive multiple gene data sets obtained via targeted PCR could provide a cost-effective solution to this problem. However, oligonucleotide primers for PCR amplification of nuclear genes across a wide range of hymenopteran species are still scarce. Here we present a suite of degenerate oligonucleotide primer pairs for PCR amplification of 154 single-copy nuclear protein-coding genes from Hymenoptera. These primers were inferred from genome sequence data from nine Hymenoptera (seven species of ants, the honeybee, and the parasitoid wasp Nasonia vitripennis). We empirically tested a randomly chosen subset of these primer pairs for amplifying target genes from six Hymenoptera, representing the families Chrysididae, Crabronidae, Gasteruptiidae, Leucospidae, Pompilidae, and Stephanidae. Based on our results, we estimate that these primers are suitable for studying a large number of nuclear genes across a wide range of apocritan Hymenoptera (i.e., all hymenopterans with a wasp-waist) and of aculeate Hymenoptera in particular (i.e., apocritan wasps with stingers). The amplified nucleotide sequences are (a) with high probability from single-copy genes, (b) easily generated at low financial costs, especially when compared to phylogenomic approaches, (c) easily sequenced by means of an additionally provided set of sequencing primers, and (d) suitable to address a wide range of phylogenetic questions and to aid rapid species identification via barcoding, as many amplicons contain both exonic and fast-evolving intronic nucleotides.
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影响因子：
4.1
作者：
Desjardins, Christopher A.;Regier, Jerome C.;Mitter, Charles
通讯作者：
Mitter, Charles
影响因子：
3.7
作者：
Feau N;Decourcelle T;Husson C;Desprez-Loustau ML;Dutech C
通讯作者：
Dutech C
DOI：
10.1073/pnas.91.21.9911
发表时间：
1994-10-11
影响因子：
11.1
作者：
DOWTON, M;AUSTIN, AD
通讯作者：
AUSTIN, AD
影响因子：
5.4
作者：
Peters RS;Meyer B;Krogmann L;Borner J;Meusemann K;Schütte K;Niehuis O;Misof B
通讯作者：
Misof B
影响因子：
4.1
作者：
Sharanowski, Barbara J.;Robbertse, Barbara;Sharkey, Michael J.
通讯作者：
Sharkey, Michael J.