Information on the following vectors is provided in three formats. A jpeg image of the vector map [map], the vector sequence in FASTA format [FASTA], and the vector sequence in VectorNTI molecule document format [VectorNTI]. The VectorNTI molecule document allows visualization of the map in conjunction with restriction analysis when viewed with VectorNTI's free document viewer, which is available from Informax Inc.
Our standard transcription vectors were produced by Jeff Essner by combining pBluescript (Stratagene) with a cDNA encoding the Xenopus B-glogin 3' UTR (a gift of H. Joseph Yost's lab). pBluescript provides the T7 and T3 promoter sites for in vitro mRNA transcription, and the B-globin 3' UTR stabilizes the mRNA. The five vectors have the 3' UTR inserted in various positions within the multiple cloning site of pBluescript or in opposing orientations.
Our fish vectors (FV) contain the carp ß-actin proximal enhancer/promoter element along with exon-1 (non-coding) and intron1 of the same gene. These are flanked by the Chinook salmon growth hormone 3'UTR including the poly(A) signal. The various vectors contain different restriction sites behind the ß-actin intron 1 as well as completing flanking the expression cassette. These vectors were described in Caldovic, L. & Hackett, P.B. Jr. (1995) Mol Mar Biotechnol 4(1): 51-61.
The Sleeping Beauty transposon system is a binary system that consists of a source of the Sleeping Beauty transposase and a DNA transposon. The DNA transposon can consist of apparently any DNA sequence so long as it as flanked by the inverted repeats (IRs) recognized by the transposase. Since each inverted repeat contains a pair of direct repeats (DRs) they are often referred to as IR/DRs. The reconstruction of the Sleeping Beauty transposon system from molecularly extinct transposons from many fish species is discussed in Ivics, Z., Hackett, P.B. Jr., Plasterk, R.H., & Izsvak, Z. (1997) Cell 91: 501-510.
Dicistronic vectors containing the EMCV internal ribosome entry site were constructed to study the regulation of protein synthesis during zebrafish development. These vectors were used to demonstrate for the first time that cap-dependent and cap-independent translation are indeed active during zebrafish embryogenesis (Fahrenkrug, S.C., K. Clark, and P.B. Hackett (1999). Mar. Biotech. 1: 552-561). Co-expression of effector (1st cistron) and reporter (2nd cistron) gene-products from a single mRNA provides a method for mosaic analysis of gene-function in zebrafish.
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