WormBase Tree Display for Construct: WBCnstr00010226
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WBCnstr00010226 | Other_name | Expr1336_Ex | |
---|---|---|---|
Summary | [unc-104::gfp] | ||
Driven_by_gene | WBGene00006831 | ||
Gene | WBGene00006831 | ||
Fusion_reporter | GFP | ||
Type_of_construct | Translational_fusion | ||
Construction_summary | [unc-104::gfp] translational fusion, a 2.3 kb SphI-AatII unc-104 fragment (including exons 510) from cosmid C52E12 was cloned into pGEM-7Zf vector and named Part II. A 3.0 kb fragment representing the 3 end of the unc-104 gene (exons 1022) was PCR-amplified from the cDNA clone yk16g10 using the primers 5'-tatgctcaacaagaacttc-3 and 5'-caactgcagtgaagcag-caattgaagatg-3 . This 3.0 kb fragment was shuttled through the pGEM-T vector (Promega) and was named Part III. Ligating the 2.3 kb SphI-AatII from Part II with the 2.7 kb AatII-PstI fragment from Part III generated a 5.0 kb fragment. This 5.0 kb SphI-PstI fragment was cloned into the SphI-PstI site in pPD95.77 and named Part II III. A 6.2kb SphI-SphI fragment from cosmid C47A5 was characterized and found to overlap a fragment of cosmid C52E12. This SphI-SphI fragment extended from 3.7 kb upstream of the first exon to the intron between exons 4 and 5 of unc-104. The plasmid clone of UNC-104::GFP was constructed by inserting the 6.2 kb SphI-SphI fragment into Part II III in the correct orientation. --precise ends. | ||
Clone | C52E12 | ||
pPD95.77 | |||
C47A5 | |||
yk16g10 | |||
Used_for | Transgene_construct | WBTransgene00027470 | |
Reference | WBPaper00004704 |