Cloning of spinach glycolate oxidase cDNA and its construction into yeast expression vector

Zhi Bin Liu; Guo Guang Chen; Shuang Li; Pin Ting He; Ping Wei; Ping Kai Ouyang

Cloning of spinach glycolate oxidase cDNA and its construction into yeast expression vector

Zhi Bin Liu, Guo Guang Chen, Shuang Li, Pin Ting He, Ping Wei, Ping Kai Ouyang

生物与制药工程学院

科研成果: 期刊稿件 › 文章 › 同行评审

摘要

METHOD: Using spinach to extract total RNA, the spinach GO cDNA was amplified by RT-PCR. This cDNA was inserted into cloning vector pMD-T and sequenced. The cDNA was then cloned into pPIC3.5K (P. Pastoris expression vector) by SnaBI/NotI double digestion. RESULT: Screening by PCR, we got the recombinant plasmid pPIC3.5K-GO which was confirmed by SnaBI/NotI restriction enzyme analysis. Sequencing showed that this cDNA was spinach spinach glycolate oxidase cDNA. The SnaBI/NotI double digestion of pPIC3.5K-GO ensured that the construction is correct. CONCLUSION: Obtained spinach glycolate oxidase cDNA and its recombinant plasmid pPIC3.5K-GO which will be useful for further expression study.

源语言	英语
页（从-至）	81-84
页数	4
期刊	Journal of China Pharmaceutical University
卷	34
期	1
出版状态	已出版 - 2月 2003

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@article{f68752b481684cda8f2d31229ad1166e,

title = "Cloning of spinach glycolate oxidase cDNA and its construction into yeast expression vector",

abstract = "METHOD: Using spinach to extract total RNA, the spinach GO cDNA was amplified by RT-PCR. This cDNA was inserted into cloning vector pMD-T and sequenced. The cDNA was then cloned into pPIC3.5K (P. Pastoris expression vector) by SnaBI/NotI double digestion. RESULT: Screening by PCR, we got the recombinant plasmid pPIC3.5K-GO which was confirmed by SnaBI/NotI restriction enzyme analysis. Sequencing showed that this cDNA was spinach spinach glycolate oxidase cDNA. The SnaBI/NotI double digestion of pPIC3.5K-GO ensured that the construction is correct. CONCLUSION: Obtained spinach glycolate oxidase cDNA and its recombinant plasmid pPIC3.5K-GO which will be useful for further expression study.",

keywords = "Cloning, Spinach glycolate oxidase, Yeast expression vector",

author = "Liu, {Zhi Bin} and Chen, {Guo Guang} and Shuang Li and He, {Pin Ting} and Ping Wei and Ouyang, {Ping Kai}",

year = "2003",

month = feb,

language = "英语",

volume = "34",

pages = "81--84",

journal = "Journal of China Pharmaceutical University",

issn = "1000-5048",

publisher = "China Pharmaceutical University",

number = "1",

}

TY - JOUR

T1 - Cloning of spinach glycolate oxidase cDNA and its construction into yeast expression vector

AU - Liu, Zhi Bin

AU - Chen, Guo Guang

AU - Li, Shuang

AU - He, Pin Ting

AU - Wei, Ping

AU - Ouyang, Ping Kai

PY - 2003/2

Y1 - 2003/2

N2 - METHOD: Using spinach to extract total RNA, the spinach GO cDNA was amplified by RT-PCR. This cDNA was inserted into cloning vector pMD-T and sequenced. The cDNA was then cloned into pPIC3.5K (P. Pastoris expression vector) by SnaBI/NotI double digestion. RESULT: Screening by PCR, we got the recombinant plasmid pPIC3.5K-GO which was confirmed by SnaBI/NotI restriction enzyme analysis. Sequencing showed that this cDNA was spinach spinach glycolate oxidase cDNA. The SnaBI/NotI double digestion of pPIC3.5K-GO ensured that the construction is correct. CONCLUSION: Obtained spinach glycolate oxidase cDNA and its recombinant plasmid pPIC3.5K-GO which will be useful for further expression study.

AB - METHOD: Using spinach to extract total RNA, the spinach GO cDNA was amplified by RT-PCR. This cDNA was inserted into cloning vector pMD-T and sequenced. The cDNA was then cloned into pPIC3.5K (P. Pastoris expression vector) by SnaBI/NotI double digestion. RESULT: Screening by PCR, we got the recombinant plasmid pPIC3.5K-GO which was confirmed by SnaBI/NotI restriction enzyme analysis. Sequencing showed that this cDNA was spinach spinach glycolate oxidase cDNA. The SnaBI/NotI double digestion of pPIC3.5K-GO ensured that the construction is correct. CONCLUSION: Obtained spinach glycolate oxidase cDNA and its recombinant plasmid pPIC3.5K-GO which will be useful for further expression study.

KW - Cloning

KW - Spinach glycolate oxidase

KW - Yeast expression vector

UR - http://www.scopus.com/inward/record.url?scp=30044431943&partnerID=8YFLogxK

M3 - 文章

AN - SCOPUS:30044431943

SN - 1000-5048

VL - 34

SP - 81

EP - 84

JO - Journal of China Pharmaceutical University

JF - Journal of China Pharmaceutical University

IS - 1

ER -

Cloning of spinach glycolate oxidase cDNA and its construction into yeast expression vector

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