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真菌快速缺失生产<em>农杆菌</em>介入OSCAR删除构造的转换

JoVE Journal
Genetics

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JoVE Journal Genetics

Rapid Deletion Production in Fungi via Agrobacterium Mediated Transformation of OSCAR Deletion Constructs

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真菌快速缺失生产<em>农杆菌</em>介入OSCAR删除构造的转换

DOI:

10.3791/55239-v

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11:24 min

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June 12, 2017

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Scott E. Gold, Zahi Paz, María D. García-Pedrajas, Anthony E. Glenn

¹Toxicology and Mycotoxin Research Unit,NPRC, USDA-ARS, ²Hazera Seeds LTD, Brurim, ³Instituto de Hortofruticultura Subtropical y Mediterránea “La Mayora” – Universidad de Málaga – Consejo Superior de Investigaciones Científicas (IHSM-UMA-CSIC), Estación Experimental “La Mayora”

Chapters

00:05Title
01:02Primer Design for PCR Amplification of Gene Flanks
02:49Production of OSCAR Constructs
05:48Agrobacterium tumefaciens Mediated Transformation (ATMT) of Fungi
07:45Deletion Mutant Identification by PCR
08:53Results: The OSCAR Method Generates a Gene Deletion Plasmid for Fungi in a Single Reaction
10:23Conclusion

Summary

Automatic Translation

通过同源重组产生的基因缺失突变体是基因功能研究的黄金标准。描述了用于快速产生缺失构建体的OSCAR（农杆菌 - 重组 - 质粒的一步构建）方法。 土壤杆菌介导的真菌转化如下。最后，提出了基于真菌转化体基因缺失的基于PCR的确认方法。

Tags

Rapid Deletion Production Fungi Agrobacterium OSCAR Deletion Constructs Homologous Recombination Functional Genetics Gene Deletion Mutants DelsGate System Primer Design PCR Deletion Confirmation

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