Table 1

Levels of 10A1 MLV and A-MLV entry supported by human PiT2 and derived truncation mutants^a.

Construct^b

No. (%) of cells infected^c

A-MLV

10A1 MLV

Expt 1^d

Expt 2^d

Expt 4

Expt 1^d

Expt 2^d

Expt 3^d

Expt 4

Expt 5

PiT2 (pOJ74)

100 ±22

100 ± 8

100 ±12

100 ± 5

100 ± 8

100 ± 9

100 ±11

100 ± 9

PiT2ΔL₁₈₃-V₄₈₃

25 ± 1

13 ± 2

ND

10 ± 1

12 ± 4

24 ± 3

ND

25 ± 3

PiT2ΔR₂₅₄-V₄₈₃

ND^e

ND

4 ± >1

ND

ND

ND

1 ± >1

1 ± >1

Empty vector^f

<0.0008

<0.002

<0.01

<0.002

<0.001

<0.001

<0.007

<0.003

^aThe experimental setup is described in the text. A-MLV and 10A1 MLV vector pseudotypes were tested on the same precipitates made from CsCl-purified plasmids in experiment 4. In experiment 5, Qiagen maxiprep-purified plasmids were used for preparing precipitates and only the 10A1 MLV vector pseudotype was tested.

^bReceptor and mutant receptor sequences were cloned into pcDNA1A^RtkpA.

^cThe data are averages of three independent transfections ± SEM. The average number of blue cells per three 60-mm-diameter dishes transfected with a plasmid encoding PiT2 was assigned a value of 100% (57,000, 90,000, and 32,000 blue cells per dish for A-MLV in experiment 4 and for 10A1 MLV in experiments 4 and 5, respectively).

^dData are from Bøttger and Pedersen 2004 [31]; please see article for details.

^eND, not determined.

^fValues are based on the detection limit of 1 blue cell per three 60-mm-diameter dishes.

Bøttger and Pedersen BMC Biochemistry 2011 12:21   doi:10.1186/1471-2091-12-21