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CDC42-Interacting Protein 4 Gene Is Down Trans-Regulated by HBV DNA polymerase Trans Activated Protein 1.

Lun Y, Xu C, Chi Q, Wang X, Sui W, Jiang S - Iran. J. Public Health (2014)

Bottom Line: The cDNAs were sequenced and analyzed with BLAST search against the sequences in GenBank.HBVDNAPTP1 may be involved in the positive regulation on the initiation of monocyte apoptosis.The result contribute to reveal the HBVDNAPTP1 biological functions and provide new evidences for further exploration of the regulatory mechanism of HBVDNAPTP1.

View Article: PubMed Central - PubMed

Affiliation: 1. Liaoning Provincial University Key Laboratory of Biophysics, College of Medicine, Dalian University Dalian, China.

ABSTRACT

Background: Hepatitis B Virus (HBV) DNA polymerase transactivated protein 1 (HBVDNAPTP1) is a novel protein transactivated by HBV DNA polymerase, screened by suppression subtractive hybridization technique (GenBank accession no: AY450389). The biological function of HBVDNAPTP1 was investigated in this study.

Methods: We constructed a vector pcDNA3.1 (-)/myc-His A-HBVDNAPTP1 and used it to transfect acute monocytic leukemia cell line THP-1. HBVDNAPTP1 expression was detected by western blot analysis in the cells. A cDNA library of genes transactivated by HBVDNAPTP1 in THP-1 cells was made in pGEM-T Easy using suppression subtractive hybridization (SSH). The cDNAs were sequenced and analyzed with BLAST search against the sequences in GenBank.

Results: Some sequences, such as CIP4, might be involved in apoptosis development. mRNA and protein expression of CIP4 was identified by Real time RT-PCR and western blot in THP-1 cells. HBVDNAPTP1 could down-regulate the expression of CIP4 at both transcription and translation levels.

Conclusion: HBVDNAPTP1 may be involved in the positive regulation on the initiation of monocyte apoptosis. The result contribute to reveal the HBVDNAPTP1 biological functions and provide new evidences for further exploration of the regulatory mechanism of HBVDNAPTP1.

No MeSH data available.


Related in: MedlinePlus

Analysis of subtracted cDNA library (Reduction of G3PDH abundance showed high subtraction efficiency). Unsubtracted secondary PCR product (lanes 1-4) and subtracted (lanes 5-8). DNA Marker DL2,000 (lane M). 18 cycles (lanes 1, 5). 23 cycles (lanes 2, 6). 28 cycles (lanes 3, 7). 33 cycles (lanes 4, 8)
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Figure 3: Analysis of subtracted cDNA library (Reduction of G3PDH abundance showed high subtraction efficiency). Unsubtracted secondary PCR product (lanes 1-4) and subtracted (lanes 5-8). DNA Marker DL2,000 (lane M). 18 cycles (lanes 1, 5). 23 cycles (lanes 2, 6). 28 cycles (lanes 3, 7). 33 cycles (lanes 4, 8)

Mentions: Subtraction efficiency analysis showed that PCR products of the housekeeping gene G3PDH in unsubtracted library were obviously visible after 18 cycles, however, 28 cycles were required in the subtracted one (Fig.3), indicating that the abundance of non-differentially expressed gene was effectively reduced and the subtraction method had high subtraction efficiency. Using SSH, a total of one hundred positive clones were obtained.


CDC42-Interacting Protein 4 Gene Is Down Trans-Regulated by HBV DNA polymerase Trans Activated Protein 1.

Lun Y, Xu C, Chi Q, Wang X, Sui W, Jiang S - Iran. J. Public Health (2014)

Analysis of subtracted cDNA library (Reduction of G3PDH abundance showed high subtraction efficiency). Unsubtracted secondary PCR product (lanes 1-4) and subtracted (lanes 5-8). DNA Marker DL2,000 (lane M). 18 cycles (lanes 1, 5). 23 cycles (lanes 2, 6). 28 cycles (lanes 3, 7). 33 cycles (lanes 4, 8)
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4419165&req=5

Figure 3: Analysis of subtracted cDNA library (Reduction of G3PDH abundance showed high subtraction efficiency). Unsubtracted secondary PCR product (lanes 1-4) and subtracted (lanes 5-8). DNA Marker DL2,000 (lane M). 18 cycles (lanes 1, 5). 23 cycles (lanes 2, 6). 28 cycles (lanes 3, 7). 33 cycles (lanes 4, 8)
Mentions: Subtraction efficiency analysis showed that PCR products of the housekeeping gene G3PDH in unsubtracted library were obviously visible after 18 cycles, however, 28 cycles were required in the subtracted one (Fig.3), indicating that the abundance of non-differentially expressed gene was effectively reduced and the subtraction method had high subtraction efficiency. Using SSH, a total of one hundred positive clones were obtained.

Bottom Line: The cDNAs were sequenced and analyzed with BLAST search against the sequences in GenBank.HBVDNAPTP1 may be involved in the positive regulation on the initiation of monocyte apoptosis.The result contribute to reveal the HBVDNAPTP1 biological functions and provide new evidences for further exploration of the regulatory mechanism of HBVDNAPTP1.

View Article: PubMed Central - PubMed

Affiliation: 1. Liaoning Provincial University Key Laboratory of Biophysics, College of Medicine, Dalian University Dalian, China.

ABSTRACT

Background: Hepatitis B Virus (HBV) DNA polymerase transactivated protein 1 (HBVDNAPTP1) is a novel protein transactivated by HBV DNA polymerase, screened by suppression subtractive hybridization technique (GenBank accession no: AY450389). The biological function of HBVDNAPTP1 was investigated in this study.

Methods: We constructed a vector pcDNA3.1 (-)/myc-His A-HBVDNAPTP1 and used it to transfect acute monocytic leukemia cell line THP-1. HBVDNAPTP1 expression was detected by western blot analysis in the cells. A cDNA library of genes transactivated by HBVDNAPTP1 in THP-1 cells was made in pGEM-T Easy using suppression subtractive hybridization (SSH). The cDNAs were sequenced and analyzed with BLAST search against the sequences in GenBank.

Results: Some sequences, such as CIP4, might be involved in apoptosis development. mRNA and protein expression of CIP4 was identified by Real time RT-PCR and western blot in THP-1 cells. HBVDNAPTP1 could down-regulate the expression of CIP4 at both transcription and translation levels.

Conclusion: HBVDNAPTP1 may be involved in the positive regulation on the initiation of monocyte apoptosis. The result contribute to reveal the HBVDNAPTP1 biological functions and provide new evidences for further exploration of the regulatory mechanism of HBVDNAPTP1.

No MeSH data available.


Related in: MedlinePlus