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纯化无 DNA 酶活性的粗糙脉孢菌碱性磷酸酶,用于分子生物学。

Purification of Neurospora crassa alkaline phosphatase without DNAse activity for use in molecular biology.

机构信息

Departamento de Genética, FMRP-USP, Av. Bandeirantes 3900, 14049-900, Ribeirão Preto, SP, Brazil.

出版信息

World J Microbiol Biotechnol. 1995 Sep;11(5):505-7. doi: 10.1007/BF00286362.

DOI:10.1007/BF00286362
PMID:24414902
Abstract

Alkaline phosphatase, excreted by Neurospora crassa preg (c) and purified to apparent homogeneity by 7.5% PAGE, did not show DNAase activity and removed the terminal 5'-phosphate group from plasmid Bluescript M13(+) linearized with EcoRI. The preg (c) strain may therefore replace other sources of alkaline phosphatase for use in dephosphorylating linearized plasmidial DNA in cloning experiments.

摘要

碱性磷酸酶由粗糙脉孢菌 preg (c) 分泌,并通过 7.5%PAGE 纯化至明显均一性,不显示 DNAase 活性,并从用 EcoRI 线性化的质粒 Bluescript M13(+) 中去除 5'-末端磷酸基团。因此,preg (c) 菌株可能替代其他来源的碱性磷酸酶,用于在克隆实验中使线性化的质粒 DNA 去磷酸化。

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Insertional mutagenesis in Neurospora crassa: cloning and molecular analysis of the preg+ gene controlling the activity of the transcriptional activator NUC-1.粗糙脉孢菌中的插入诱变:控制转录激活因子NUC-1活性的preg+基因的克隆与分子分析
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Genetic control of alkaline phosphatase synthesis in Neurospora: the use of partial diploids in dominance studies.
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