Characterization of CIb1 Gene Promoter from Silkworm, Bombyx mori

2007 ◽  
Vol 62 (11-12) ◽  
pp. 875-880 ◽  
Author(s):  
Qiao-Ling Zhao ◽  
Xing-Jia Shen ◽  
Liang-Jun Zhu ◽  
Yong-Zhu Yi ◽  
Shun-Ming Tang ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Bombyx Mori ◽  
Transcription Initiation ◽  
Genomic Dna ◽  
Initiation Site ◽  
Transcription Initiation Site ◽  
Transcription Activity ◽  
Silkworm Bombyx Mori

The hemolymph chymotrypsin inhibitor b1 (CIb1) of silkworm, Bombyx mori, plays an important role in innate immunity. In order to study its encoding gene CIb1, five heterogeneous promoter fragments of 844 bp, 682 bp, 516 bp, 312 bp and 82 bp in length were cloned from genomic DNA of the p50 silkworm strain. Characterization of the CIb1 promoter was performed in vitro using the firefly luciferase gene as reporter. The results showed that CIb1 promoter fragments have transcription activities in the B. mori ovary-derived BmN cell line. The 82 bp fragment (-72 to +10 nt) containing the eukaryotic core promoter elements revealed a basic transcription activity. The Bm1 element, upstream the transcription initiation site, showed a positive regulation function to the CIb1 promoter. CIb1 promoter-like fragments from the genomic DNA of the tetra hybrid silkworm Suju\Minghu provided a natural deletion model for the study of the CIb1 promoter. In vitro analysis indicated that the 132 bp fragment from -517 nt to -386 nt upstream of the transcription initiation site strongly suppressed the transcription activity of the CIb1 promoter, suggesting that the 132 bp fragment harbours strong negative cis-acting elements. Infection of Bombyx mori nucleopolyhedrovirus (BmNPV) increased the activity of the CIb1 promoter, having provided another evidence to the function of CIb1 in the innate immunity of silkworm.

Identification of a new promoter upstream of the murine dihydrofolate reductase gene

1989 ◽  
Vol 9 (10) ◽  
pp. 4568-4570
Author(s):  
L J Schilling ◽  
P J Farnham
Keyword(s):  
Dihydrofolate Reductase ◽  
Transcription Initiation ◽  
Initiation Site ◽  
Transcription Initiation Site ◽  
Deletion Mapping ◽  
Reductase Gene ◽  
Bona Fide ◽  
Translational Start

In vitro reactions identified a transcription initiation site located 740 nucleotides upstream of the dihydrofolate reductase translational start. Transcription from this site proceeded in the direction opposite to that of dihydrofolate reductase mRNA. Deletion mapping indicated that this new promoter can be separated from the dihydrofolate reductase promoter and that separation increased transcription at -740. Transcripts that initiate at -740 were also detected in cellular RNA, indicating that this is a bona fide transcription initiation site in vivo.

Functional characterization of short-type peptidoglycan recognition proteins (PGRPs) from silkworm Bombyx mori in innate immunity

2019 ◽  
Vol 95 ◽  
pp. 59-67 ◽  
Author(s):  
Ruo-Nan Zhang ◽  
Cai-ting Li ◽  
Fei-fei Ren ◽  
Ming-Qiang Ye ◽  
Xiao-juan Deng ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Bombyx Mori ◽  
Functional Characterization ◽  
Silkworm Bombyx Mori ◽  
Short Type

scholarly journals Characterization of the Promoter of PRS1 inSaccharomyces cerevisiae Identifies Three Regions Potentially Involved in Control of Expression

2001 ◽  
Vol 183 (2) ◽  
pp. 795-799 ◽  
Author(s):  
Yolanda Hernando ◽  
Andrew T. Carter ◽  
Stefan Sickinger ◽  
Michael Schweizer
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Transcription Initiation ◽  
Initiation Site ◽  
Transcription Initiation Site ◽  
Galactosidase Activity ◽  
Shift Analysis ◽  
Gel Shift ◽  
Control Of Expression

ABSTRACT The transcription initiation site of the Saccharomyces cerevisiae PRS1 gene was mapped at −179 bp. Measurement of β-galactosidase activity of the successively deleted PRS1promoter linked to lacZ and integrated at theura3 locus defined three DNA regions involved in the control of PRS1 expression. Gel shift analysis confirmed the data.

scholarly journals Cellular factor YY1 downregulates the human papillomavirus 16 E6/E7 promoter, P97, in vivo and in vitro from a negative element overlapping the transcription-initiation site

2009 ◽  
Vol 90 (10) ◽  
pp. 2402-2412 ◽  
Author(s):  
Michael J. Lace ◽  
Yasushi Yamakawa ◽  
Masato Ushikai ◽  
James R. Anson ◽  
Thomas H. Haugen ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Transcription Initiation ◽  
Regulatory Region ◽  
Initiation Site ◽  
Transcription Initiation Site ◽  
Wild Type ◽  
Hpv 16 ◽  
Human Papillomavirus 16

Cellular factors that bind to cis sequences in the human papillomavirus 16 (HPV-16) upstream regulatory region (URR) positively and negatively regulate the viral E6 and E7 oncogene promoter, P97. DNase I footprinting has revealed the binding of cellular proteins to two previously undetected cis elements overlapping and 3′ of the transcription-initiation site of the P97 promoter. Mutations within homologous motifs found in both of these cis elements abolished their negative function in vivo and the binding of the same cellular complex in vitro. This factor was identified as YY1 by complex mobility and binding specificity in comparison with vaccinia virus-expressed, purified recombinant YY1 protein and by antigenic reactivity with YY1 antisera. Cis mutations in the ‘initiator’ YY1 site activated the P97 promoter in vivo and in vitro. P97 was also activated threefold in vitro by depletion of endogenous YY1 with wild-type, but not mutant, YY1 oligonucleotides from the IgH kappa E3′ enhancer. Furthermore, increasing concentrations of exogenous, purified recombinant YY1 repressed wild-type P97 transcript levels by up to threefold, but did not influence the P97 promoter mutated in the ‘initiator’ YY1 site. Thus, the promoter-proximal YY1 site was not necessary for correct transcription initiation at the P97 promoter, but was found to be required for downregulation of P97 transcription in vivo and in vitro. In contrast to other viral and cellular promoters, where YY1 is thought to function as a positive transcription-‘initiator’ factor, HPV-16 P97 transcription is downregulated by YY1 from a critical motif overlapping the transcription start site.

scholarly journals Isolation and characterization of the androgen-dependent mouse cysteine-rich secretory protein-1 (CRISP-1) gene

1997 ◽  
Vol 321 (2) ◽  
pp. 325-332 ◽  
Author(s):  
Uta SCHWIDETZKY ◽  
Wolf-Dieter SCHLEUNING ◽  
Bernard HAENDLER
Keyword(s):  
Transcription Initiation ◽  
Initiation Site ◽  
Secretory Protein ◽  
Mobility Shift ◽  
Isolation And Characterization ◽  
Eukaryotic Genes ◽  
Mobility Shift Assay ◽  
Responsive Element

In mice, cysteine-rich secretory protein-1 (CRISP-1) is mainly found in the epididymis and also, to a lesser extent, in the salivary gland of males, where androgens control its expression. We have now isolated and characterized overlapping phage clones covering the entire length of the CRISP-1gene. DNA sequencing revealed that the gene is organized into eight exons, ranging between 55 and 748 bp in size, and seven introns. All exonŐintron junctions conformed to the GT/AG rule established for eukaryotic genes. The intron length, as determined by PCR, varied between 1.05 and 4.0 kb so that the CRISP-1gene spans over 20 kb of the mouse genome. The transcription-initiation site was determined by primer extension and localized at the expected distance downstream of a consensus TATA box. Approximately 3.7 kb of the CRISP-1promoter region were isolated and sequenced, and several stretches fitting the androgen-responsive element consensus were found. Those that most resembled the consensus were analysed by electrophoretic mobility-shift assay and found to form specific complexes with the liganded androgen receptor in vitro, but with different affinities. Putative binding elements for the transcription factors Oct, GATA, PEA3, CF1, AP-1 and AP-3 were also found in the promoter region.

scholarly journals Identification of a new promoter upstream of the murine dihydrofolate reductase gene.

1989 ◽  
Vol 9 (10) ◽  
pp. 4568-4570 ◽  
Author(s):  
L J Schilling ◽  
P J Farnham
Keyword(s):  
Dihydrofolate Reductase ◽  
Transcription Initiation ◽  
Initiation Site ◽  
Transcription Initiation Site ◽  
Deletion Mapping ◽  
Reductase Gene ◽  
Bona Fide ◽  
Translational Start

In vitro reactions identified a transcription initiation site located 740 nucleotides upstream of the dihydrofolate reductase translational start. Transcription from this site proceeded in the direction opposite to that of dihydrofolate reductase mRNA. Deletion mapping indicated that this new promoter can be separated from the dihydrofolate reductase promoter and that separation increased transcription at -740. Transcripts that initiate at -740 were also detected in cellular RNA, indicating that this is a bona fide transcription initiation site in vivo.

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