Table 1

Propagation of MMTV and MPMV transfer vectors RNA by homologous and heterologous proteins.

Titers (CFU/ml)^{a, b}

% EGFP

Positive Cells^c

Transfer Vector

Description of the Transfer Vector

MMTV Protein (JA10)

MPMV Protein (TR301)

MMTV Protein (JA10)

MPMV Protein (TR301)

pDA024

Chimeric LTR, 5' region upto 400 bp of MMTV Gag, and SV-hyg^r

3,676 ± 196

< 1

-

-

pSS013

Chimeric LTR, 5' region upto 400 bp of MMTV Gag, and SV-EGFP

-

-

19

< 1

pND015

Same as DA024 but the putative MMTV ψ has been replaced with that of MPMV (5' UTR + 282 bp of Gag)

2

8

-

-

pND016

Same as DA024 but the putative MMTV ψ and PBS have been replaced with that of MPMV

< 1

7

-

-

pND017

Control MMTV vector in which putative MMTV ψ has been cloned back after creating NotI site at PBS/UTR junction

187 ± 33

ND

-

-

pND018

Control MMTV vector in which putative MMTV ψ and PBS have been cloned back after creating NotI site at U5/PBS junction

30 ± 4

ND

-

-

pKAL011

MPMV 5' region upto 282 bp Gag and SV-hyg^r

< 1

36,373 ± 3,972

-

-

pSS015

Same as KAL011 but has SV-EGFP instead of SV-hyg^r

-

-

< 1

41

pND011

Same as KAL011 but MPMV ψ has been replaced with that of MMTV (5' UTR + 400 bp of Gag)

< 1

3

-

-

pND012

Same as KAL011 but MPMV ψ and PBS have been replaced with that of MMTV

< 1

23

-

-

pND013

Control MPMV vector in which MPMV ψ has been cloned back after creating NotI site at PBS/UTR junction

ND

2813 ± 99

-

-

pND014

Control MPMV vector in which MPMV ψ and PBS have been cloned back after creating NotI site at U5/PBS junction

ND

1660 ± 111

-

-

pTR174

Control vector containing SV-hyg^r cassette and SIV 3'LTR as poly(A)

< 1

< 1

-

-

pAG001

Control vector containing SV-hyg^r cassette and FIV 3'LTR as poly(A)

< 1

< 1

-

-

Mock

No DNA (control)

<1

< 1

< 1

< 1

^a CFU/ml, colony forming units per milliliter of non-concentrated supernatant from the transfected cultures. ^b Each value represents a mean of three experiments performed in duplicates. ^c Percentage of EGFP positive cells represent the phenotypic analysis of the transduced target cells. Single cell suspensions of the infected HeLa CD4+ cells were prepared, and 10,000 events (cells) per group were counted and analyzed using Becton Dickinson FACS. < 1 indicates < 1 Hyg^r colonies in the target cells or < 1 EGFP positive cells out of 10,000 events (cells) analyzed.

Al Dhaheri et al. Retrovirology 2009 6:66   doi:10.1186/1742-4690-6-66