PMID- 2288881
OWN - NLM
STAT- MEDLINE
DCOM- 19910402
LR  - 20141120
IS  - 1044-9523 (Print)
IS  - 1044-9523 (Linking)
VI  - 1
IP  - 9
DP  - 1990 Sep
TI  - Expression of the human retinoblastoma gene product pp110RB in insect cells using 
      the baculovirus system.
PG  - 429-37
AB  - The product of the retinoblastoma susceptibility gene (RB) was overproduced in 
      cultured insect cells using the baculovirus expression system. Upon insertion of 
      the cloned human RB complementary DNA sequence into the viral genome downstream 
      of the promoter of the polyhedrin gene, full-length RB protein with an apparent 
      molecular weight of 110,000 was expressed in the insect cells. This protein was 
      found to be phosphorylated, located in the nuclei of the infected cells, and 
      immunologically indistinguishable from pp110RB of human cells as assayed by 
      several anti-RB antibodies. Following cell disruption and a one-step 
      immunoaffinity chromatographic purification, 6-12 mg of soluble pp110RB with 
      approximately 95% purity were obtained per liter of infected suspension culture. 
      Characterization of the two known biochemical properties of RB protein showed 
      that this purified protein from insect cells behaved similarly to the authentic 
      human pp110RB. First, it bound to DNA, and second, it could form a specific 
      complex with SV40 T antigen in vitro. Prompt translocation of the protein from 
      cytoplasm to nucleus after microinjection further indicated that the purified RB 
      protein may be active. The availability of soluble, intact, and presumably active 
      pp110RB in large quantity represents a significant advance for studying the 
      biochemical and biophysical properties of the RB gene product as well as its 
      potential biological function in cancer suppression.
FAU - Wang, N P
AU  - Wang NP
AD  - Department of Pathology, University of California, San Diego, La Jolla 92093.
FAU - Qian, Y W
AU  - Qian YW
FAU - Chung, A E
AU  - Chung AE
FAU - Lee, W H
AU  - Lee WH
FAU - Lee, E Y
AU  - Lee EY
LA  - eng
GR  - CA 49649/CA/NCI NIH HHS/United States
GR  - EY05758/EY/NEI NIH HHS/United States
GR  - EY57980/EY/NEI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Cell Growth Differ
JT  - Cell growth & differentiation : the molecular biology journal of the American 
      Association for Cancer Research
JID - 9100024
RN  - 0 (Antigens, Polyomavirus Transforming)
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Macromolecular Substances)
RN  - 0 (Nuclear Proteins)
RN  - 0 (Phosphoproteins)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Retinoblastoma Protein)
SB  - IM
MH  - Animals
MH  - Antigens, Polyomavirus Transforming/metabolism
MH  - Baculoviridae/genetics
MH  - Base Sequence
MH  - Blotting, Western
MH  - Cell Compartmentation
MH  - Cell Nucleus/metabolism
MH  - Cells, Cultured
MH  - DNA-Binding Proteins/genetics/metabolism
MH  - Gene Expression
MH  - Genetic Vectors
MH  - In Vitro Techniques
MH  - Macromolecular Substances
MH  - Molecular Sequence Data
MH  - Moths
MH  - Nuclear Proteins/metabolism
MH  - Phosphoproteins/metabolism
MH  - Precipitin Tests
MH  - Recombinant Proteins/genetics/metabolism
MH  - Retinoblastoma Protein/*genetics
EDAT- 1990/09/01 00:00
MHDA- 1990/09/01 00:01
CRDT- 1990/09/01 00:00
PHST- 1990/09/01 00:00 [pubmed]
PHST- 1990/09/01 00:01 [medline]
PHST- 1990/09/01 00:00 [entrez]
PST - ppublish
SO  - Cell Growth Differ. 1990 Sep;1(9):429-37.