pRSVlacZII (ATCC® 77129)

Applications: contains sequence beta-galactosidase beta-D-galactosidaseexpression vectorin another host, produces protein beta-galactosidase beta-D-galactosidasereporter constructshuttle vectorvector permitting production of single-stranded DNAvector permitting visual detection of activity in colonies  /  Depositors: LA Culp

Designations pRSVlacZII
Depositors LA Culp
Biosafety Level 1
Host
Distribution host: Escherichia coli DH5alpha
Vector Information
Size (kb): 9.0000000000000000
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: pRSVlacZII (phagemid)
Promoters: Promoter RSV LTR
Construction: pRc/RSV, pM-MuLV-SV-lacZ
Marker(s):G418R,ampR,neoR
Construct size (kb): 8.899999618530273
Features: marker(s): ampR, neoR, G418R
promoter: RSV LTR
replicon: pMB1, M13
terminator: bGH polyadenylation
enhancer: RSV LTR
Applications
contains sequence beta-galactosidase beta-D-galactosidase
expression vector
in another host, produces protein beta-galactosidase beta-D-galactosidase
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--4.4, 3.4, 1.2; HindIII--9.0; BglII--9.0, KpnI--4.5 (doublet); XbaI--9.0.
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 beta-galactosidase 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 3.9 kb HindIII/BamHI fragment from pM-MuLV-SV-lacZ into pRc/RSV.
Media ATCC® 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

Sanes JR, et al. Use of a recombinant retrovirus to study post-implantation cell lineage in mouse embryos. EMBO J. 5: 3133-3142, 1986. PubMed: 3102226

Lloyd A Culp, personal communication

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