| *** The vector sequence has now been corrected as of 7/13/04. The WAP/KCR/bGHpA cassette was inverted in the pBluescript vector ***
WKbpA: WAP5' cloned into KbpA vector (originally cloned by S. Chua, Baylor College of Medicine)
WAP5' = -944 to +33 rat Whey Acidic Protein (WAP) promoter
KbpA vector:
KCR = 640 bp portion of rabbit B-globin gene containing 18 bp of exon II, 572 bp of intron 2, and 50 bp of exon II
- NOTES:
•Having an intron 5' of your gene greatly increases mammary gland-specific expression, even of large cDNAs
(we found expression of 3.3 kb cDNA).
•The 5' intron works much better than a 3' intron, probably by disguising the cDNA as a terminal exon instead
of an internal exon (large terminal exons occur in nature, while internal exons are almost always quite small).
bpA = bovine Growth Hormone poly A signal
The WKbpA construct was subcloned into pBluescript II, which contains BssHII sites flanking the MCS. These are used to cut out the whole transgene for microinjection into embryos. This final construct is called WKbpAII.
Clone your gene of interest into the EcoRI site at 3870, downstream of the KCR element.
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