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Species: Xenopus laevis
Genetic Insert: dnWnt-11
Vector Backbone Description: Backbone Marker:Available at Addgene (#15030); Backbone Size:3300; Vector Backbone:pSP64T; Vector Types:Xenopus Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:10769246
Comments: A C-terminally truncated dominant-negative Xwnt11 construct (comprising amino acids 1-282) was generated according to Hoppler
et al. (1996). PCR was carried out using pGEM-X9 (Ku and Melton, 1993) as a template and the primers 5′-CCCCCTCGAGAGTACCAATGGCTCCGACCCG-3′ and 5′-TTGGAGATCTTTCAGCAGTAGTCAGGGGAAC-3′. The PCR product obtained was cloned into the XhoI and BglII sites of pSP64TXB.
For injection, linearize with BamH1 and transcribe with SP6.
Proper citation: RRID:Addgene_16857 Copy
Species: Xenopus laevis
Genetic Insert: Engrailed-2
Vector Backbone Description: Backbone Marker:Stratagene; Backbone Size:3000; Vector Backbone:pBluescript KS; Vector Types:Xenopus Probe; Bacterial Resistance:Ampicillin
Defining Citation: PMID:1679005
Comments: In situ probe cut with Xba1, transcribe with T3.
Proper citation: RRID:Addgene_16950 Copy
Species: Xenopus laevis
Genetic Insert: Xwnt-5A
Vector Backbone Description: Backbone Size:3000; Vector Backbone:pSP64T; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:8275867
Comments: Insert: cDNA from Melton lambda gt10 oocyte library.
Transcription: linearize with Xba1 and transcribe with SP6.
Plasmid History:
Constructed by Randall Moon on date of 9/15/90.
Notes for Use:
EcoR1 insert of lambda G6 of 2/11/90. Contains significant 5' UTR and is less translatable in vitro per ng RNA than Xwnt-5A Hind III/SP64T. However, the type of activity is the same in both constructs.
Citation: Moon et al., Development 119, 97 (1993)
Proper citation: RRID:Addgene_16985 Copy
Species: Xenopus laevis
Genetic Insert: banded hedgehog
Vector Backbone Description: Backbone Marker:Stratagene; Backbone Size:3000; Vector Backbone:pBluescript KS; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: Xenopus Hedgehog. Antisense are generated as listed:
Xenopus banded hedgehog: Linearize clone XIL in Bluescript KS with BamH1 and transcribe with T3. This is a ~2.5 Kb EcoR1 insert.
Xenopus sonic hedgehog: Linearize clone X32 in Bluescript KS with Bam H1 and transcribe with T3. This is a ~2.5 Kb EcoR1 insert.
Xenopus cephalic hedgehog:Linearize clone X30 in pT7TS with Sal 1 and transcribe with T7.
Proper citation: RRID:Addgene_16984 Copy
Species: Xenopus laevis
Genetic Insert: Goosecoid
Vector Backbone Description: Backbone Size:3000; Vector Backbone:SP64T; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: Insert: coding region of Xenopus goosecoid in SP64T in sense (#9) & antisense (#2) orientation.
Transcription: linearize with EcoR1 and transcribe with SP6.
Plasmid History:
Constructed by Jan Christian by PCR.
Goosecoid cloned and characterized by lab of E. DeRobertis, UCLA.
Proper citation: RRID:Addgene_16979 Copy
Species: Xenopus laevis
Genetic Insert: Xbra
Vector Backbone Description: Backbone Marker:Promega; Backbone Size:2500; Vector Backbone:psp73; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: Insert: full length Xbra from Jim Smith.
Antisense: linearize with EcoRV, transcribe with T7.
Plasmid History:
Constructed by Jan Christian.
Proper citation: RRID:Addgene_16978 Copy
Species: Xenopus laevis
Genetic Insert: noggin PCR clone
Vector Backbone Description: Backbone Size:4120; Vector Backbone:pCSKA; Vector Types:Xenopus Expression; Bacterial Resistance:Ampicillin
Comments: Insert: noggin PCR clone (coding region). In reverse orientation compared to CSKA promoter.
Plasmid History:
Constructed by Jan Christian.
Isolated and characterize by W. Smith and R. Harland.
Proper citation: RRID:Addgene_16998 Copy
Species: Xenopus laevis
Genetic Insert: noggin
Vector Backbone Description: Backbone Size:3000; Vector Backbone:SP64T; Vector Types:Xenopus Expression; Bacterial Resistance:Ampicillin
Comments: Insert: noggin PCR clone (coding region) in SP64T.
Transcription: linearize with EcoR1 and transcribe with SP6.
Plasmid History:
Constructed by Jan Christian.
Noggin isolated and chacterized by W. Smith and R. Harland.
Proper citation: RRID:Addgene_16997 Copy
Species: Xenopus laevis
Genetic Insert: Xbhh-C
Vector Backbone Description: Backbone Size:3000; Vector Backbone:pT7TS; Vector Types:Xenopus Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:7671801
Comments: Expression ConstructXenopus Banded Hedgehog.
(a.k.a. Xbhh; hh1)
Insert: PCR of coding region, Genbank accession number U26404.
Antisense RNA: For in situ we use clone XIL in Bluescript KS. Linearize with Bam H1 and transcribe with T3.
Sense RNA: Linearize with Bam H1 and transcribe with T7.
References:
Ekker et al., Development 121, 2337-2347 (1995)
Lai et al., Development 121, 2349-2360 (1995)
Subclone Xbhh-N in pT7Ts.
Encodes amino terminal product with autoproteolytic cleavage. Linearize with BamH1 and transcribe with T7.
Subclone Xbhh-dominant negative-C in pT7TS:
Encodes a leader sequence, a few amino acids amino terminal to the autoproteolytic site, and the carboxy-terminal domain. Linearize with BamH1 and transcribe with T7.
Proper citation: RRID:Addgene_16995 Copy
Species: Xenopus laevis
Genetic Insert: Xbhh
Vector Backbone Description: Backbone Size:3000; Vector Backbone:pT7TS; Vector Types:Xenopus Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:7671801
Comments: Expression Construct Xenopus Banded Hedgehog (a.k.a. Xbhh; hh1)
Insert: PCR of coding region, Genbank accession number U26404.
Antisense RNA: For in situ we use clone XIL in Bluescript KS. Linearize with Bam H1 and transcribe with T3.
Sense RNA: Linearize with Bam H1 and transcribe with T7.
References:
Ekker et al., Development 121, 2337-2347 (1995)
Lai et al., Development 121, 2349-2360 (1995)
Proper citation: RRID:Addgene_16993 Copy
Species: Xenopus laevis
Genetic Insert: C-cadherin
Vector Backbone Description: Backbone Size:0; Vector Backbone:pSP36T; Vector Types:Xenopus Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:7556920
Comments: Linearize with EcoRI and transcribe with SP6.
A cDNA encoding Xenopus C-cadherin was cloned by screening a Xenopus lambda gt10 oocyte cDNA library. The cDNA was instered into pSP36T for transcription using the SP6 promoter.
Proper citation: RRID:Addgene_17023 Copy
Species: Xenopus laevis
Genetic Insert: dominant N-cadherin
Vector Backbone Description: Backbone Size:3000; Vector Backbone:pT7TS; Vector Types:Xenopus Expression; Bacterial Resistance:Ampicillin
Comments: Full-length N-cadherin was digested with BamHI to remove nt 90-2004, deleting the extracellular domain while
keeping the signal sequence (identical to preparation by C. Kintner, 1992, Cell 69:225-236, who provided the cadherin).
Linearize and transcribe with T7.
Proper citation: RRID:Addgene_17025 Copy
Species: Xenopus laevis
Genetic Insert: beta-globin 5' UTR
Vector Backbone Description: Backbone Size:0; Vector Backbone:pCD-PS; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Comments: Description: This is an old but serviceable SV40 expression construct (pCD-PS) modified to give the Xenopus beta-globin untranslated regions and multiple cloning sites from pT7TS vector (Addgene plasmid 17091).
Reference: The vector was made by Julia Yang-Snyder.
Proper citation: RRID:Addgene_17120 Copy
Species: Xenopus laevis
Genetic Insert: Sizzled
Vector Backbone Description: Backbone Size:4100; Vector Backbone:pCS2+; Vector Types:Xenopus Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:9428410
Comments: This encodes a secreted
antagonist of Xwnt-8.
The entire open reading frame of sizzled was amplified by low-cycle number PCR using Pfu polymerase and cloned as a
BamHI-XhoI fragment into pCS2+ .
Proper citation: RRID:Addgene_16688 Copy
Species: Xenopus laevis
Genetic Insert: APC
Vector Backbone Description: Backbone Size:4350; Vector Backbone:pCS2+MT; Vector Types:Xenopus Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:9015311
Comments: Insert: Full length Xenopus APC with an amino terminal c-myc epitope in the vector pCS2+MT.
Insert size: ~8.5kb. NcoI site introduced at 5' end. Linearize with Not1 and transcribe with SP6.
Proper citation: RRID:Addgene_16686 Copy
Species: Xenopus laevis
Genetic Insert: PABPC1 (M161A)
Vector Backbone Description: Vector Backbone:pGEM_11Zf; Vector Types:in vitro transcription template; Bacterial Resistance:Ampicillin
Defining Citation: PMID:34213414
Comments: The absolute length of the poly(A) region may not be accurate. The poly(A) region may not be completely maintained during amplification from bacteria. Single clones need to be screened to verify poly(A) length.
Proper citation: RRID:Addgene_174349 Copy
Species: Xenopus laevis
Genetic Insert: ePAB
Vector Backbone Description: Vector Backbone:pcDNA5; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:34213414
Proper citation: RRID:Addgene_174359 Copy
Species: Xenopus laevis
Genetic Insert: Smurf1
Vector Backbone Description: Vector Backbone:pCS2+; Vector Types:Xenopus expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:10458166
Proper citation: RRID:Addgene_117244 Copy
Species: Xenopus laevis
Genetic Insert: Tbx5-EnR
Vector Backbone Description: Vector Backbone:pCS2+; Vector Types:Xenopus expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:10079235
Proper citation: RRID:Addgene_117249 Copy
Species: Xenopus laevis
Genetic Insert: BRAT
Vector Backbone Description: Vector Backbone:pCS2+; Vector Types:Xenopus expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:9165117
Proper citation: RRID:Addgene_117251 Copy
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