【原创】发现一个一篇文章的一个错误。
丁香园论坛
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文献的作者：Suzuki J, Fukuda M, Kawata S, Maruoka M, Kubo Y, Takeya T, Shishido T.
题目：A rapid protein expression and purification system using Chinese hamster ovary cells expressing retrovirus receptor.
期刊：J Biotechnol. 2006 Dec 1;126(4):463-74.
PMID: 16766074
2.1. Cloning of EcoR
An ecotropic retrovirus receptor (EcoR) was amplified
by polymerase chain reaction (PCR) using cDNA
derived from NIH3T3 cells as a template, the forward
primer 5-gctaccgtggactctgctgtg-3, and the reverse
primer 5-ggcgaattctcatttgcactggtccaagttgc-3. cDNA
was prepared as described below in Section 2.4. The
PCR product was digested with BamHI and EcoRI,
inserted into the BamHI and EcoRI sites of pcDNA3.1
(Invitrogen), and verified by sequencing. The product
was then digested with BamHI and NotI to excise the
EcoR fragment, which was subcloned into pCX4 bsr
(Akagi et al., 2003) (a gift from Tsuyoshi Akagi, Osaka
Bioscience Institute), yielding pCX4 bsr EcoR.
粗粗的阅读没问题。
仔细的研究就有问题。
1，先从PUBMED上找到 ecotropic retrovirus receptor (EcoR)的 CDNA序列，序列号是D67087。
2，在看他设计的引物。the forward primer 5-gctaccgtggactctgctgtg-3,
就有毛病了。（你感兴趣的话，想想为什么不对！）
the reverse primer 5-ggcgaattctcatttgcactggtccaagttgc-3.
应该是对的。
3.在看他的酶切位点。晕倒！the forward primer 根本没有 BamHI位点。
题目：A rapid protein expression and purification system using Chinese hamster ovary cells expressing retrovirus receptor.
期刊：J Biotechnol. 2006 Dec 1;126(4):463-74.
PMID: 16766074
2.1. Cloning of EcoR
An ecotropic retrovirus receptor (EcoR) was amplified
by polymerase chain reaction (PCR) using cDNA
derived from NIH3T3 cells as a template, the forward
primer 5-gctaccgtggactctgctgtg-3, and the reverse
primer 5-ggcgaattctcatttgcactggtccaagttgc-3. cDNA
was prepared as described below in Section 2.4. The
PCR product was digested with BamHI and EcoRI,
inserted into the BamHI and EcoRI sites of pcDNA3.1
(Invitrogen), and verified by sequencing. The product
was then digested with BamHI and NotI to excise the
EcoR fragment, which was subcloned into pCX4 bsr
(Akagi et al., 2003) (a gift from Tsuyoshi Akagi, Osaka
Bioscience Institute), yielding pCX4 bsr EcoR.
粗粗的阅读没问题。
仔细的研究就有问题。
1，先从PUBMED上找到 ecotropic retrovirus receptor (EcoR)的 CDNA序列，序列号是D67087。
2，在看他设计的引物。the forward primer 5-gctaccgtggactctgctgtg-3,
就有毛病了。（你感兴趣的话，想想为什么不对！）
the reverse primer 5-ggcgaattctcatttgcactggtccaagttgc-3.
应该是对的。
3.在看他的酶切位点。晕倒！the forward primer 根本没有 BamHI位点。
