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Volumn 282, Issue 5387, 1998, Pages 284-287

Alterations of the PPP2R1B gene in human lung and colon cancer

Author keywords

[No Author keywords available]

Indexed keywords

ARTICLE; CANCER GENETICS; CHROMOSOME 11Q; COLON CANCER; GENE LOCATION; GENE MUTATION; GENE SEQUENCE; HUMAN; LUNG CANCER; NUCLEOTIDE SEQUENCE; PRIORITY JOURNAL; TUMOR SUPPRESSOR GENE;

EID: 0032500792     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.282.5387.284     Document Type: Article
Times cited : (332)

References (33)
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    • note
    • Amplification was carried out with a 5′ RACE Kit (Boehringer Mannheim) and oligonucleotide primers 5′-TCACTTCGGGTCCTTTCTACTCCA-3′ and 5′-CTTCATTGCGGAGCTCGTCGA-3′. Amplification conditions were as follows: 95°C for 11 s. 59°C for 11 s, and 72°C for 15 s, for 33 cycles.
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    • note
    • RNA was prepared from lung cancer-derived and matched normal (lymphoblastoid) cell lines with the RNA/DNA STAT60 Kit (Tel-Test, Friendswood, TX). Primary tumor samples were obtained from the Cooperative Tissue Network (Birmingham, AL), and cancer cell lines were obtained from the American Type Culture Collection (ATCC) and A. Gazdar's lab. Randomly primed cDNA was synthesized with Superscript II (Gibco-BRL) and PCR was performed with the Expand Long Template PCR System (Boehringer Mannheim). In certain cases, nested PCR was performed with 20-fold-diluted amplification products from the initial PCR amplification. Primer pairs were as follows: 5′-GGTGACC-AGCAGCAGGAG-3′ and 5′-GCTTGGATGAGATC-TTGAAGGPCR-3′, 5′-GCGCATCAGAGCTCGGGACCG-3′ and 5′-CCATTCTTTCTCCACCCAGTTAAGAAC-3′. Amplification products were separated on 0.8% agarose gels, DNA bands were isolated with the QIA Quick Gel Extraction Kit (Qiagen, Santa Clarita, CA), and the DNA was sequenced with the Dye Terminator Sequencing Kit (Perkin-Elmer, Foster City, CA).
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    • Gel-purified PCR products were cloned with the TOPO-TA Cloning Kit (Invitrogen, Carlsbad, CA). DNA from a single transformed colony was isolated and sequenced by means of automated DNA sequencing.
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    • data not shown
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  • 31
    • 3643107063 scopus 로고    scopus 로고
    • note
    • Cells were lysed, proteins were separated by electrophoresis, and gels were transferred to Millipore Immobilon-P membrane. Immunodetection was carried out with goat anti-human PP2A-Aβ immunoglobulin and secondary horseradish peroxidase-conjugated donkey anti-goat immunoglobulin (Santa Cruz Biotechnology). Immunodetection was carried out with enhanced chemiluminescence (Amersham Life Sciences). Goat anti-human actin immunoglobulin was added as a control.
  • 32
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    • note
    • Cell lysates were immunoprecipitated with rabbit anti-human PP2A-C immunoglobulin (Promega) and protein C-agarose (Boehringer Mannheim). The precipitate was collected by centrifugation, and the pellet was washed with cold phosphate-buffered saline. The precipitate was then analyzed on an 8% SDS-polyacrylamide gel. Protein immunoblot detection was carried out with purified goat anti-human immunoglobulin against PP2A-Aβ and PP2A-C and secondary horseradish peroxidase-conjugated donkey anti-goat immunoglobulin (Santa Cruz Biotechnology).
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    • note
    • We thank C. Richard III, M. Mumby, S. Federov, P. Ramos, J. Korcz, D. Stickens, V. Gee, and P. Schilling for assistance and helpful discussions. Supported by grants from NIH and the U.S. Department of Energy to G.A.E. and by a Lung Cancer SPORE grant from the National Cancer Institute to J.M. and A.G. E.D.E. was a predoctoral trainee of the Medical Scientist Training Program.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.