甲型流感病毒IgG基因捕獲法檢測(cè)試劑盒
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甲型流感病毒IgG基因捕獲法檢測(cè)試劑盒
廣州健侖生物科技有限公司
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甲型流感病毒IgG基因捕獲法檢測(cè)試劑盒
基因捕獲法是利用隨機(jī)插入突變進(jìn)行基因敲除的新型方法。通?；虿东@載體還包括一個(gè)無(wú)啟動(dòng)子的報(bào)道基因，通常是neo 基因，neo 基因插入到ES 細(xì)胞染色體組中，并利用捕獲基因的轉(zhuǎn)錄調(diào)控元件實(shí)現(xiàn)表達(dá)的ES 克隆可以很容易地在含G418 的選擇培養(yǎng)基中篩選出來(lái)，從理論上講，在選擇培養(yǎng)基中存活的克隆應(yīng)該100%地含有中靶基因。中靶基因的信息可以通過(guò)篩選標(biāo)記基因側(cè)翼cDNA或染色體組序列分析來(lái)獲得。
基因捕獲法是利用隨機(jī)插入突變進(jìn)行基因敲除的新型方法。通常基因捕獲載體還包括一個(gè)無(wú)啟動(dòng)子的報(bào)道基因，通常是neo 基因，neo 基因插入到ES 細(xì)胞染色體組中，并利用捕獲基因的轉(zhuǎn)錄調(diào)控元件實(shí)現(xiàn)表達(dá)的ES 克隆可以很容易地在含G418 的選擇培養(yǎng)基中篩選出來(lái)，從理論上講，在選擇培養(yǎng)基中存活的克隆應(yīng)該100%地含有中靶基因。中靶基因的信息可以通過(guò)篩選標(biāo)記基因側(cè)翼cDNA或染色體組序列分析來(lái)獲得。
用常規(guī)方法進(jìn)行基因敲除研究需耗費(fèi)大量的時(shí)間和人力，研究者必須針對(duì)靶位點(diǎn)在染色體組文庫(kù)中篩選相關(guān)的染色體組克隆，繪制相應(yīng)的物理圖譜，構(gòu)建特異性的基因敲除載體以及篩選中靶ES 細(xì)胞等，通常一個(gè)基因剔除純合子小鼠的獲得需要一年或更長(zhǎng)的時(shí)間。面對(duì)人類基因組計(jì)劃產(chǎn)生出來(lái)的巨大的功能未知的遺傳信息，傳統(tǒng)的基因敲除方法顯得有些力不從心。因此，基因捕獲法應(yīng)運(yùn)而生，利用基因捕獲可以建立一個(gè)攜帶隨機(jī)插入突變的ES 細(xì)胞庫(kù)，節(jié)省大量篩選染色體組文庫(kù)以及構(gòu)建特異打靶載體的工作及費(fèi)用，更有效和更迅速地進(jìn)行小鼠染色體組的功能分析。
此方法的缺點(diǎn)是只能剔除在Es 細(xì)胞中表達(dá)的基因．單種的細(xì)胞類型中表達(dá)的基因數(shù)目約為I04，現(xiàn)在的基因捕獲載體從理論上來(lái)講應(yīng)能剔除所有在ES細(xì)胞表達(dá)的基因，因此，在ES 細(xì)胞中進(jìn)行基因捕獲還是大有可為的。用基因捕獲法進(jìn)行基因剔除的另一個(gè)缺點(diǎn)是無(wú)法對(duì)基因進(jìn)行精細(xì)的遺傳修飾。
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【公司名稱】 廣州健侖生物科技有限公司
【市場(chǎng)部】 楊永漢
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【騰訊 】 2042552662
【公司地址】 廣州清華科技園創(chuàng)新基地番禺石樓鎮(zhèn)創(chuàng)啟路63號(hào)二期2幢101-103室
Gene capture is a novel method of gene knockout using random insertions. Generally, the gene capture vector also includes a promoterless reporter gene, usually a neo gene. The neo gene is inserted into the genome of the ES cell and an ES clone expressing the transcriptional regulatory element of the capture gene can be easily expressed in a G418-containing The selection medium is screened, and theoretically, the clone that survives in the selection medium should contain 100% of the target gene. The target gene information can be obtained by screening the flanking cDNA or genomic sequence analysis of the marker gene.
Gene capture is a novel method of gene knockout using random insertions. Generally, the gene capture vector also includes a promoterless reporter gene, usually a neo gene. The neo gene is inserted into the genome of the ES cell and an ES clone expressing the transcriptional regulatory element of the capture gene can be easily expressed in a G418-containing The selection medium is screened, and theoretically, the clone that survives in the selection medium should contain 100% of the target gene. The target gene information can be obtained by screening the flanking cDNA or genomic sequence analysis of the marker gene.
Gene knockout studies using conventional methods take a considerable amount of time and manpower. Researchers have to screen for the relevant genomic clones in the genomic library against the target site, map the corresponding physical maps, construct specific knockout vectors, and Screening of target ES cells and the like usually takes one year or more to obtain a gene knockout homozygous mouse. In the face of huge untapped genetic information generated by the Human Genome Project, traditional methods of gene knock-outs seem a bit powerless. Therefore, the gene capture method came into being, the use of gene capture can create a library of ES cells carrying random insertion mutations, saving a large number of screening of the genome library and the construction of specific targeting vector work and costs more efficient and more prompt mouse chromosome Group functional analysis.
The disadvantage of this method is that only the genes expressed in Es cells can be excluded. The number of genes expressed in a single cell type is about 104, and now the gene capture vector is theoretically able to knock out all the genes expressed in ES cells. Therefore, gene capture in ES cells is still promising. Another drawback of gene knockout using gene trapping is the inability to fine-tune genes for genetic modification.
