pRSVADH (ATCC® 77131)

Applications: contains sequence alcohol dehydrogenaseexpression vectorin another host, produces protein alcohol dehydrogenasereporter constructshuttle vectorvector permitting production of single-stranded DNAvector permitting visual detection of activity in colonies  /  Depositors: LA Culp

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Designations pRSVADH
Depositors LA Culp
Biosafety Level 1
Host
Distribution host: Escherichia coli DH5alpha
Vector Information
Size (kb): 6.9000000953674320
DESCRIPTION OF VECTOR COMPONENT:
Name of vector: pRc/RSV
Intact vector size: 5.100
Type of vector: phagemid
Vector end: HindIII
Vector end: XbaI
Cloning sites: HindIII SpeI BstXI NotI XbaI
Polylinker sites: HindIII KpnI SacI BamHI SpeI XmaIII BstXI EcoRI PstI EcoRV
BstXI NotI XhoI SphI NsiI XbaI
Construction: pUC19, RSV, bovine growth hormone
Host range: vertebrate cells; Escherichia coli
Features (with orientation and position when available):
marker(s): ampR, neoR, G418R
promoter: RSV LTR
replicon: pMB1, M13
terminator: bGH polyadenylation
Cross references:
Vector: pRSVADH (phagemid)
Promoters: Promoter RSV LTR
Construction: pRc/RSV, pWX0008
Marker(s):G418R,ampR,neoR
Construct size (kb): 6.900000095367432
Features: marker(s): ampR, neoR, G418R
promoter: RSV LTR
replicon: pMB1, M13
terminator: bGH polyadenylation
enhancer: RSV LTR
Applications
contains sequence alcohol dehydrogenase
expression vector
in another host, produces protein alcohol dehydrogenase
reporter construct
shuttle vector
vector permitting production of single-stranded DNA
vector permitting visual detection of activity in colonies
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI--4.3, 2.6; HindIII--6.9; BglII--6.9; KpnI--6.9; XbaI--6.9; HindIII/XbaI--5.1, 1.8.
Transient expression experiments should be performed to examine the RSV promoter efficiency in different cell lines. The RSV promoter can be replaced by excising with a BglII/HindIII digest.
Shuttle reporter plasmid permitting visual detection of activity by histochemical staining.
Presence of alcohol dehydrogenase activity is used to follow cell lineage in culture or in situ.
pRSVlacZII (ATCC 77129), pRSVPAP (ATCC 77130) and pRSVADH (ATCC 77131) provide distinct color-staining reactions (aqua blue, red, and blue-black respectively) to permit simultaneous analysis of multiple lineages.
The order of the major features in pRc/RSV is: RSV LTR - MCS - bovine growth hormone polyadenylation signal - M13 ori - SV40 early promoter - neoR - SV40 polyadenylation signal - pMB1 ori - bla.
Constructed by cloning the HindIII/XbaI fragment from pWX0008 into pRc/RSV.
Media Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Lin WC, Culp LA. Selectable plasmid vectors with alternative and ultrasensitive histochemical marker genes. BioTechniques 11: 344-351, 1991. PubMed: 1931036

Shen NL, et al. Analysis of adh gene regulation in Drosophila: studies using somatic transformation. Dev. Genet. 10: 210-219, 1989. PubMed: 2500285

Lloyd A Culp, personal communication

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