• Title/Summary/Keyword: Northern blot analysis

Search Result 432, Processing Time 0.028 seconds

Cloning and Characterization of the psbEF Gene Encoding Cytochrome b-559 of the Panax ginseng Photosystem II Reaction Center

  • Lee, Won-Kyu;Park, Dae-Sung;Tae, Gun-Sik
    • BMB Reports
    • /
    • v.32 no.2
    • /
    • pp.189-195
    • /
    • 1999
  • From the Panax ginseng chloroplast, the psbE and psbF genes, encoding the $\alpha$- and $\beta$-subunits of cytochrome b-559 of the photosystem II reaction center, respectively, were cloned and characterized. The psbE and psbF genes were composed of 252 and 117 nucleotides, respectively. The deduced amino acid sequence of the $\alpha$-subunits showed 95%, 93%, and 91% homology to monocots, dicots, and liverwort, respectively, whereas the $\beta$-subunits showed approximately 98% to 95% homology to the same species. Southern blot analysis revealed that a single copy of the psbEF gene exists in the chloroplast plastid. Northern blot analysis indicated that the psbE and psbF genes are cotranscribed as a polycistron.

  • PDF

Isolation and Characterization of a cDNA for a Ribulose-1,5-Bisphosphate Carboxylase Small Subunit in Spinach

  • Jin, Yun-Hae;Park, Yang-Seo;Jeong, Ji-Na;Cho, Tae-Ju;Cho, Nam-Jeong
    • BMB Reports
    • /
    • v.30 no.3
    • /
    • pp.173-176
    • /
    • 1997
  • We isolated a cDNA clone that encodes a ribulose-1,5-bisphosphate carboxylase small subunit (rbcS) from spinach using a soybean rbcS cDNA as a probe. The small subunit consists of 180 amino acids including a transit peptide of 57 residues. Comparison of the amino acid sequence with those of other plant species shows a maximum of 70-80% identical residues. Southern blot analysis suggests the existence of multiple rbcS genes in the spinach genome. Northern blot analysis indicates that the rbcS gene is expressed predominantly in leaves and that the expression of the gene is induced by light.

  • PDF

Cloning and characterization of the psbA Gene from Panax ginseng(Characterization of the psbA Gene from P. ginseng)

  • Lee, Won-Kyu;Tae, Gun-Sik
    • Journal of Photoscience
    • /
    • v.10 no.3
    • /
    • pp.245-249
    • /
    • 2003
  • The psbA gene of photo system II was cloned and characterized from the P. ginseng chloroplast. The psbA gene is composed of 1,062 nucleotides. The overall amino acid sequence shows 99% and 98% identities to dicots and monocots of higher plants, respectively. Southern blot analysis revealed that a single copy of the psbA gene existed in the chloroplast genome. Northern blot analysis of the in vivo accumulation of the psbA transcript, after being grown under the different intensities (5%, 10%, 20%, and 100%) of daylight, indicated that the steady-state level of the psbA transcript was not significantly affected by light intensity.

  • PDF

Characterization of ORF39 from Helicoverpa armigera Single-nucleocapsid Nucleopolyhedrovirus, the Gene Containing RNA Recognition Motif

  • Xu, Hai-Jun;Liu, Yan-He;Yang, Zhang-Nv;Zhang, Chuan-Xi
    • BMB Reports
    • /
    • v.39 no.3
    • /
    • pp.263-269
    • /
    • 2006
  • In the genome of Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus, open reading frame 39 (Ha39) is the only gene predicted to encode an RNA recognition protein. Computer analysis revealed that Ha39 homologues were found in 15 NPVs, but not in GVs. Its transcripts were detected from 3 through 72 hours post infection (h p.i.) using RT-PCR and Northern blot analysis. The protein was detected in infected-cell lysates from 6 h p.i. Western blot assay of ODV and BV preparations revealed that Ha39 encodes a structural protein associated with BVs. Additionally, immunofluorescence microscopy demonstrated that the protein was present within cytoplasm in virus-infected cells, but not in the nuclear region.

Screening for Inhibitor of c-myc Expression and Identification of Isolate No.2303

  • Chung, Ji-Hyung;Yeo, Ick-Hyun;Oh, Doo-Whan;Moon, Soon-Ok
    • Journal of Microbiology and Biotechnology
    • /
    • v.5 no.5
    • /
    • pp.264-268
    • /
    • 1995
  • Sulforhodamine B(SRB) assay was performed on the human lung carcinoma, A549 cell line to screen soil microorganisms for production of anti-cancer agent. Among 4, 265 microorganisms, 45 isolates were selected for their cytotoxicity and tested for their effects on the expression of c-myc by RNA slot blot and Northern blot analysis resulting in selection of No.2303 isolate. This No.2303 was identified as Streptomyces sp. by ISP classification and the chemotaxonomic analysis method. NO.2303 inhibited the expression of cmyc in Col0320 DM and A549 cell lines. The culture extract of No. 2303 also inhibited the progression of the cell cycle of Go in NIH 313 cells, implying that the extract also inhibited the expression of c-myc in NIH 313 cell.

  • PDF

Development of Potato Virus Y Resistant Tobacco Plant by Transformation of the Untranslatable Viral Coat Protein Encoding cDNA (감자 바이러스 Y 비전이성 외피단백질 cDNA의 형질전환에 의한 바이러스 저항성 연초품종 개발)

  • 이청호;이영기;강신웅;박성원;김상석;박은경
    • Journal of the Korean Society of Tobacco Science
    • /
    • v.19 no.2
    • /
    • pp.117-123
    • /
    • 1997
  • Viral coat protein (CP) encoding cDNA with artificial start and stop codons was synthesized by reverse-transcriptase polymerase chain reaction (RT-PCR) from the Korean isolate of potato virus Y-vein nectrosis strain (pVY-VN). To make PVY CP cDNA to untranslatable form, three stop codons were inserted near the start codon by "megaprimer-PCR" method. The untranslatable CP cDNA was subcloned to plant expression vector and transferred to N. tabacum cv. NC82 by Agrobacterium-mediated transformation. Highly resistant plants to PVY infection were screened, based on symptom development after mechanical virus inoculation. By genomic PCR and Southern blot analysis, one or more copies of the untranslatable CP gene were found in all transformants. From northern blot analysis, highly resistant transgenic lines had very low level of CP transcript but susceptible lines had high level, suggesting resistance to PVY infection should be related to RNA-mediated mechanism.mechanism.

  • PDF

A cDNA Clone for the 5' Exon of Chloroplast ATP Synthase Subunit I Gene (atpF) from Broccoli (Brassica oleracea L. var. Italica) and Its Expression Pattern

  • Choo Bong Hong
    • Journal of Plant Biology
    • /
    • v.38 no.2
    • /
    • pp.137-141
    • /
    • 1995
  • We isolated a cDNA clone, BLSC1, encoding 5' exon of ATP synthase CF0 subunit I from broccoli. BLSC1 is 285 nucleotides long which consists of a 5' noncoding region of 34 nucleotides, a 5' exon of 145 nucleotides and an intron of 106 nucleotides. The 5' exon codes for 48 amino acids which reveals mostly hydrophobic. The amino acid sequence deduced from BLSC1 shares 83%, 83% and 91% identities with the genes coding for atpF from wheat, rice and spinach, respectively. Genomic Southern blot analysis for BLSC1 showed a typically strong signal for a gene located in the chloroplast genome. Northern blot analysis identified three major classes of transcripts showing strong positive signals in the leaves, but only trace amounts of the transcripts were identified in the other organs like stems, flowr buds and roots.

  • PDF

Expression of Aspergillus awamori Glucoamylase Gene in Asperillus nidulans (Aspergillus nidulans내에서 Aspergillus awamori의 Glucoamylase 유전자 발현)

  • 김석준;유준희;정구홍
    • Korean Journal of Microbiology
    • /
    • v.31 no.2
    • /
    • pp.136-140
    • /
    • 1993
  • The A. nidulans expression vector which contained trpC marker gene from A. nidulans was constructed to produce glucoamy]ase. The recombinant plasmid was introduced into auxotrophic mutant A. nidulans B17. Southern blot analysis of the genomic DNA from transformant showed that pKHG2 DNA had integrated into the A. nidulans chromosomes. Northern analysis of the total RNA from transform ant showed that mRNA of glucoamylase gene was synthesized in induction condition. Specific activity of glucoamylase was increased in transform ants. G]ucoamylase was shown to be active in non-denaturing acrylamide gel.

  • PDF

Cloning of $\alpha$-Amylase Gene from Zea mays (옥수수 $\alpha$-amylase 유전자의 클로닝)

  • 김용욱;강신혜
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.38 no.3
    • /
    • pp.275-282
    • /
    • 1993
  • The objective of this study was to clone a partial fragment of $\alpha$-amylase from Korean maize. We designed and synthesized an oligonucleotide probe and two kinds of PCR primers based on cDNA conserved region of $\alpha$-amylase sequences from other plants. Total RNA from 3-day-old maize seedling was used as template for 1st strand cDNA synthesis and RNA-DNA hybrid was used as template for polymerase chain reaction(PCR). The product of PCR was about 0.5 kb long and inserted into pUC19. We named this recombinant plasmid as pZM$\alpha$'. The cloned fragment was certified by Southern blot analysis using labeled synthetic oligonucleotide as probe.

  • PDF

The Study of X Chromosome Inactivation Mechanism in Klinefelter's Syndrome by cDNA Microarray Experiment

  • Jeong, Yu-Mi;Chung, In-Hyuk;Park, Jung Hoon;Lee, Sook-Hwan;Chung, Tae-Gyu;Kim, Yong Sung;Kim, Nam-Soon;Yoo, Hyang-Sook;Lee, Suman
    • Genomics & Informatics
    • /
    • v.2 no.1
    • /
    • pp.30-35
    • /
    • 2004
  • To investigate the XIST gene expression and its effect in a Klinefelter's patient, we used Klinefelter's syndrome (XXY) patient with azoospermia and also used a normal male (XY) and a normal female (XX) as the control, We were performed cytogenetic analysis, Y chromosomal microdeletion assay (Yq), semi-quantitative RT-PCR, and the Northern blot for Klinefelter's syndrome (KS) patient, a female and a male control, We extracted total RNA from the KS patient, and from the normal cells of the female and male control subjects using the RNA prep kit (Qiagen), cDNA microarray contained 218 human X chromosome-specific genes was fabricated. Each total RNA was reverse transcribed to the first strand cDNA and was labeled with Cy-3 and Cy-5 fluorescein, The microarray was scanned by ScanArray 4000XL system. XIST transcripts were detected from the Klinefelters patient and the female by RT-PCR and Northern blot analysis, but not from the normal male, In the cDNA microarray experiment, we found 24 genes and 14 genes are highly expressed in KS more than the normal male and females, respectively. We concluded that highly expressed genes in KS may be a resulted of the abnormal X inactivation mechanism.