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百態(tài)人生銀蟲 (小有名氣)
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[求助]
分子基因方面的問題,多謝
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今天翻譯了一篇文獻,有一段話,“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,剛開始學習。 這段話大概的意思懂了,然后有幾個點不太明白,求助一下,希望大家?guī)蛶兔Α?br /> 1."用自殺載體同源重組進行無標記的基因敲除"具體是什么意思? 2."crossover PCR technique"是什么技術(shù)? 3."The resulting amplicon was ligated to pCR725, which was subsequently transferred to C. glutamicum. "之前已經(jīng)有步驟敲除基因了,為什么還說"將擴增子連接到質(zhì)粒上,轉(zhuǎn)入菌中" 一頭霧水的,希望大家指點一下,謝謝謝謝。 |
捐助貴賓 (著名寫手)
商家已經(jīng)主動聲明此回帖可能含有宣傳內(nèi)容|
Markerless gene disruption was carried out through a two-step homologous recombination using the suicide vector pCRA725 “利用自殺載體pCRA725,進行兩步式基因同源重組,從而實現(xiàn)無痕基因敲除” crossover PCR technique PCR(polymerase chain reaction) technique 實際上是一種“基因擴增技術(shù)”。從字面理解就是“聚合酶連反應(yīng)”技術(shù) 前面加上“crossover”交叉,也就是“聚合酶連反應(yīng)”技術(shù) The resulting amplicon was ligated to pCR725, which was subsequently transferred to C. glutamicum. 將產(chǎn)生的擴增產(chǎn)物捆綁在pCR725(自殺載體)上,然后將其轉(zhuǎn)移到C. glutamicum |
銀蟲 (小有名氣)
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中文我翻譯的也差不多,字面意思一樣。就是不太明白它的具體方法,求指點。 還有問題2,前面文中的“聚合酶鏈式反應(yīng)”直接說“by PCR”,但這個是“by crossover PCR”,所以我覺得這不是普通的pcr,可能是其他的? 發(fā)自小木蟲Android客戶端 |
捐助貴賓 (著名寫手)
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