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[求助]
分子基因方面的问题，多谢
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今天翻译了一篇文献，有一段话，“For overexpression of the cgR_2128 gene in C. glutamicum, the cgR_2128 gene fragment amplified by PCR with primer 1 and 2 (Supplementary Table S1) was inserted into the NdeI site of a pCRB214 plasmid that is a derivative of an Escherichia coli–C. glutamicum shuttle vector pCRB22 [18] and that contains an expression cassette composed of a tac promoter, an NdeI site and an rrnB T1 terminator. C. glutamicum cells were transformed by electroporation [19]. Markerless gene disruption was carried out through a two-step homologous recombination using the suicide vector pCRA725 [8]. For disruption of cgR_2128, the nucleotide fragments (ca. 0.9 kb) of 50- and 30-regions of cgR_2128 amplified by PCR with primer set 3–4 and primer set 5–6, respectively (Table S1) were fused bycrossover PCR technique. The resulting amplicon was ligated to pCR725, which was subsequently transferred to C. glutamicum. Gene disruption was confirmed by PCR and DNA sequencing.” 我是分子方面的新人，刚开始学习。 这段话大概的意思懂了，然后有几个点不太明白，求助一下，希望大家帮帮忙。 1."用自杀载体同源重组进行无标记的基因敲除"具体是什么意思？ 2."crossover PCR technique"是什么技术？ 3."The resulting amplicon was ligated to pCR725, which was subsequently transferred to C. glutamicum. "之前已经有步骤敲除基因了，为什么还说"将扩增子连接到质粒上，转入菌中" 一头雾水的，希望大家指点一下，谢谢谢谢。 |
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武汉一心一译 
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百态人生: 金币+15, 翻译EPI+1, ★有帮助 2016-07-03 19:17:56
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Markerless gene disruption was carried out through a two-step homologous recombination using the suicide vector pCRA725 “利用自杀载体pCRA725，进行两步式基因同源重组，从而实现无痕基因敲除” crossover PCR technique PCR（polymerase chain reaction） technique 实际上是一种“基因扩增技术”。从字面理解就是“聚合酶连反应”技术 前面加上“crossover”交叉，也就是“聚合酶连反应”技术 The resulting amplicon was ligated to pCR725, which was subsequently transferred to C. glutamicum. 将产生的扩增产物捆绑在pCR725(自杀载体)上，然后将其转移到C. glutamicum |
2楼2016-07-03 13:05:57
武汉一心一译
捐助贵宾 (著名写手)
- 翻译EPI: 502
- 应助: 8 (幼儿园)
- 金币: 2283.1
- 散金: 5914
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- 在线: 321.9小时
- 虫号: 3587652
- 注册: 2014-12-10
- 性别: GG
- 专业: 高分子合成化学
4楼2016-07-03 22:25:21