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Tissue-Specific Split sfGFP System for Streamlined Expression of GFP Tagged Proteins in the Germline.

G3 (Bethesda). 2019; 
HefelAdam,SmolikoveS
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Catalog Antibody Primary antibodies used were rabbit anti-SYP-4 (1:500), mouse anti- 166 FLAG (1:500; Sigma) and rabbit anti-OLLAS (1:1000; Genscript #A01658). Get A Quote

摘要

Identifying protein localization is a useful tool in analyzing protein function. Using GFP-fusion tags, researchers can study the function of endogenous proteins in living tissue. However, these tags are considerably large, making them difficult to insert, and they can potentially affect the normal function of these proteins. To improve on these drawbacks, we have adopted the split sfGFP system for studying the localization of proteins in the germline. This system divides the "super folder" GFP into 2 fragments, allowing researchers to use CRISPR/Cas9 to tag proteins more easily with the smaller subunit, while constitutively expressing the larger subunit from another locus. These two parts are ab... More

关键词

AKIR-1,CRISPR,GFP1-10,GFP11,RPA-1,SYP-4,germline,meiosis,sfGFP,split