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BAC selection of the large white butterfly, Pieris brassicae (Lepidoptera: Pieridae) containing orthologs of the silkworm, Bombyx mori (Lepidoptera: Bombycidae)
Applied Entomology and Zoology ( IF 1.3 ) Pub Date : 2020-01-01 , DOI: 10.1007/s13355-019-00665-7
Mizuki Ohno , Toshiaki Fujimoto , Yota Naito , Akiya Jouraku , Yuji Yasukochi , Ken Sahara

We constructed a bacterial artificial chromosome (BAC) library from the large white butterfly, Pieris brassicae (L.) (Lepidoptera: Pieridae), which composed of 15,360 clones with an average insert size of 103 kb. We designed 39 degenerate oligonucleotide primer pairs and cloned the PCR products for sequencing. From these plus pierisin data, we designed single tagged sequence primers and selected 40 BAC clones using a 3-step PCR protocol. Additional 40 BACs were sequenced to improve identification of P. brassicae BACs carrying orthologs of the silkworm, Bombyx mori (L.) (Lepidoptera: Bombycidae). Among 34 BAC clones containing multiple B. mori orthologs, sequences in 27 BACs correspond to a block (less than 1 Mb) of every B. mori chromosome. While orthologs in 5 BACs showed collinearity to each B. mori chromosome with a distance (1.4–6.8 Mb), orthologs in two BACs corresponded to genes in more than two different B. mori chromosomes.

中文翻译:

BAC 选择含有蚕的直系同源物 Bombyx mori (Lepidoptera: Bombycidae) 的大白蝶粉蝶 (Lepidoptera: Pieridae) (Lepidoptera: Pieridae)

我们从大白蝶粉蝶 (L.) (鳞翅目: 粉蝶科) 构建了一个细菌人工染色体 (BAC) 文库,它由 15,360 个克隆组成,平均插入大小为 103 kb。我们设计了 39 个简并寡核苷酸引物对,并克隆了 PCR 产物进行测序。从这些加皮瑞辛数据中,我们设计了单标签序列引物,并使用 3 步 PCR 方案选择了 40 个 BAC 克隆。对另外 40 个 BAC 进行测序,以提高对携带家蚕 Bombyx mori (L.)(鳞翅目:Bombycidae)直系同源物的 P. brasicae BAC 的鉴定。在包含多个家蚕直向同源物的 34 个 BAC 克隆中,27 个 BAC 中的序列对应于每个家蚕染色体的一个块(小于 1 Mb)。虽然 5 个 BAC 中的直系同源物与每个家蚕染色体共线性,但距离 (1.4–6.8 Mb),
更新日期:2020-01-01
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