Gene Expression Profiling via Multigene Concatemers.
Gene Expression Profiling via Multigene Concatemers.
复制标题
通过多基因串联体进行基因表达谱分析
DOI：
10.1371/journal.pone.0015711
复制
发表时间：
2011-01-18
期刊：
影响因子：
3.7
通讯作者：
Xia Y
中科院分区：
文献类型：
--
作者：
Jin K;Zheng X;Xia Y
我们建立了一种新的方法，通过多基因串联体基因表达谱（MgC-GEP），同时研究多基因表达模式。该方法包括以下步骤：（1）使用含有接头的特异性反向引物获得cDNA。(2)在聚合酶链反应（PCR）的最初1-3个循环期间，使用含有衔接子的特异性正向和反向引物扩增含有在两端具有消化位点的通用衔接子的产物。(3)在随后的4-28个循环中，通用衔接子用作引物以产生产物。(4)将产物消化并连接以产生多联体。(5)将多联体克隆到载体中并测序。然后，确定每个基因标签的出现。为了验证MgC-GEP的有效性，我们利用MgC-GEP结合实时荧光定量RT-PCR技术对弱酸诱导的酿酒酵母中的20个基因进行了分析。与real-time RT-PCR和基因芯片分析的结果相比，MgC-GEP可以同时精确地检测多个基因的转录水平。重要的是，MgC-GEP是一种具有成本效益的策略，可以在大多数实验室中广泛使用，而无需特定设备。MgC-GEP是一个潜在的强大的工具，多基因表达谱，特别是中等通量分析。
We established a novel method, Gene Expression Profiling via Multigene Concatemers (MgC-GEP), to study multigene expression patterns simultaneously. This method consists of the following steps: (1) cDNA was obtained using specific reverse primers containing an adaptor. (2) During the initial 1–3 cycles of polymerase chain reaction (PCR), the products containing universal adaptors with digestion sites at both termini were amplified using specific forward and reverse primers containing the adaptors. (3) In the subsequent 4–28 cycles, the universal adaptors were used as primers to yield products. (4) The products were digested and ligated to produce concatemers. (5) The concatemers were cloned into the vector and sequenced. Then, the occurrence of each gene tag was determined. To validate MgC-GEP, we analyzed 20 genes in Saccharomyces cerevisiae induced by weak acid using MgC-GEP combined with real-time reverse transcription (RT)-PCR. Compared with the results of real-time RT-PCR and the previous reports of microarray analysis, MgC-GEP can precisely determine the transcript levels of multigenes simultaneously. Importantly, MgC-GEP is a cost effective strategy that can be widely used in most laboratories without specific equipment. MgC-GEP is a potentially powerful tool for multigene expression profiling, particularly for moderate-throughput analysis.
登录
查看更多内容
影响因子：
48
作者：
Matsumura, Hideo;Bin Nasir, Khairun Hisam;Terauchi, Ryohei
通讯作者：
Terauchi, Ryohei
影响因子：
56.9
作者：
Golub, TR;Slonim, DK;Lander, ES
通讯作者：
Lander, ES
DOI：
10.1073/pnas.96.16.9212
发表时间：
1999-08-03
影响因子：
11.1
作者：
Perou, CM;Jeffrey, SS;Botstein, D
通讯作者：
Botstein, D
影响因子：
64.8
作者：
Sasaki, T;Matsumoto, T;Gojohori, T
通讯作者：
Gojohori, T
影响因子：
64.8
作者：
Alizadeh, AA;Eisen, MB;Staudt, LM
通讯作者：
Staudt, LM