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公司名称: Takara Bio
产品编号: 1235A
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CRISPR-PCS Protocol for Chromosome Splitting and Splitting Event Detection in Saccharomyces cerevisiae
Author:
Date:
2017-05-20
[Abstract]  Chromosome engineering is an important technology with applications in basic biology and biotechnology. Chromosome splitting technology called PCS (PCR-mediated Chromosome Splitting) has already been developed as a fundamental chromosome engineering technology in the budding yeast. However, the splitting efficiency of PCS technology is not high enough to achieve multiple splitting at a time. This protocol describes a procedure for achieving simultaneous and multiple chromosome splits in the budding yeast Saccharomyces cerevisiae by a new technology called CRISPR-PCS. At least four ... [摘要]  染色体工程是应用于基础生物学和生物技术的重要技术。染色体分裂技术称为PCS(PCR介导的染色体分裂)已经被开发为发芽酵母中的基本染色体工程技术。然而,PCS技术的分割效率不够高,不能一次实现多次分割。该协议描述了通过称为CRISPR-PCS的新技术在芽状酵母酿酒酵母中实现同时和多个染色体分裂的过程。基因组中至少四个独立的位点可以通过一次转化来分裂。与常规PCS技术相比,获得多重分裂酵母菌株的总时间和劳动力大大降低。

背景 能够快速有效地操纵多个遗传基因座或染色体区域的染色体工程技术变得越来越重要。这些技术为阐明染色体和基因组功能提供了有力的手段。此外,它可以用于通过创建广泛的遗传变体来繁殖有用的菌株。在芽殖酵母酿酒酵母(Saccharomyces ...

Construction of Glycine Oxidase Mutant Libraries by Random Mutagenesis, Site Directed Mutagenesis and DNA Shuffling
Author:
Date:
2014-10-05
[Abstract]  Glyphosate, a broad spectrum herbicide widely used in agriculture all over the world, inhibits 5-enolpyruvylshikimate-3-phosphate synthase in the shikimate pathway, and glycine oxidase (GO) has been reported to be able to catalyze the oxidative deamination of various amines and cleave the C-N bond in glyphosate (Pedotti et al., 2009). Here, in an effort to improve the catalytic activity of the glycine oxidase that was cloned from a glyphosate-degrading marine strain of Bacillus cereus (BceGO), we used a bacteriophage T7 lysis-based method for high-throughput screening of ... [摘要]  草甘膦,广泛应用于全世界农业的广谱除草剂在莽草酸途径中抑制5-烯醇丙酮酸莽草酸-3-磷酸合酶,并且已经报道甘氨酸氧化酶(GO)能够催化各种胺的氧化脱氨基, 切割草甘膦中的CN键(Pedotti等人,2009)。 在这里,为了改善从草甘膦降解的蜡状芽孢杆菌菌株(BceGO)克隆的甘氨酸氧化酶的催化活性,我们使用基于噬菌体T7裂解的方法, 通过筛选氧化酶活性并通过定向进化改造编码BceGO的基因。

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