• Title/Summary/Keyword: DNA-level

Search Result 2,021, Processing Time 0.024 seconds

Immunopreventive Effects against Murine H22 Hepatocellular Carcinoma in vivo by a DNA Vaccine Targeting a Gastrin-Releasing Peptide

  • Meko'o, Jean Louis Didier;Xing, Yun;Zhang, Huiyong;Lu, Yong;Wu, Jie;Cao, Rongyue
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.15 no.20
    • /
    • pp.9039-9043
    • /
    • 2014
  • There is a continuing need for innovative alternative therapies for liver cancer. DNA vaccines for hormone/growth factor immune deprivation represent a feasible and attractive approach for cancer treatment. We reported a preventive effect of a DNA vaccine based on six copies of the B cell epitope GRP18-27 with optimized adjuvants against H22 hepatocarcinoma. Vaccination with pCR3.1-VS-HSP65-TP-GRP6-M2 (vaccine) elicited much higher level of anti-GRP antibodies and proved efficacious in preventing growth of transplanted hepatocarcinoma cells. The tumor size and weight were significantly lower (p<0.05) in the vaccine subgroup than in the control pCR3.1-VS-TP-HSP65-TP-GRP6, pCR3.1-VS-TP-HSP65-TP-M2 or saline subgroups. In addition, significant reduction of tumor-induced angiogenesis associated with intradermal tumors of H22 cells was observed. These potent effects may open ways towards the development of new immunotherapeutic approaches in the treatment of liver cancer.

Sequence Similarity of Nuclear 18S rDNA from Morphologically Different Blades of the Seaweed Porphyra pseudolinearis (Rhodophyta) (긴잎돌김 Porphyra pseudolinearis의 엽체형간 18S rDNA 염기서열 상동성)

  • Jin Long-Guo;KIM Young-Dae;KIM Myung-Sook;JIN Hyung-Joo;CHO Ji-Young;CHOI Jae-Suk;HONG Yong-Ki;KIM Hyung Geun
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.33 no.6
    • /
    • pp.496-500
    • /
    • 2000
  • Partial fragments of nuclear 185 rDNAs from morphologically wide and narrow thalli of the seaweed Porphyra pseudolineazis were amplified and sequenced to compare their DNA homology. Both sequences of 311 base pairs showed $100{\%}$ identical each other. They showed $97.7{\%}$ similarity with a wild strain collected at Sodol in Kangwondo, and $99.4{\%}$ similarity with the GenBank accession number AB013185 of the Japanese P. pseudolinearis. Thus the morphological difference of wide and narrow blades might not be a classification criterion for the sub-species level of P. pseudolinearis.

  • PDF

DNA Toposiomerase I Inhibitor by Streptomyces sp. 7489 (방선균주 7489가 생산하는 DNA Topoisomerase I 저해제에 관한 연구)

  • Lee, Dong-Sun;Ha, Sang-Chul;Lee, Sang-Yong;Kim, Jong-Guk;Hong, Soon-Duck
    • Microbiology and Biotechnology Letters
    • /
    • v.24 no.1
    • /
    • pp.101-104
    • /
    • 1996
  • During the screening of inhibitor of DNA topoisomerase I from microbial secondary metabolites, Streptomyces melanosporofaciens 7489 which was capable of producing high level of inhibitor was selected from soil. The active compound (7489-1) was purified from the culture broth by solvent extraction, silica gel column chromatography and HPLC. The inhibitor was identified as dibutyl phthalate by spectroscopic methods of UV, $^{1}H$-NMR, $^{13}C$-NMR, DEPT and EI-MS. 7489-1 showed a strong inhibitory activity against topoisomerase I with 10 ${\mu}$M of $IC_{50}$ value.

  • PDF

Chinese Holstein Cattle Shows a Genetic Contribution from Native Asian Cattle Breeds: A Study of Shared Haplotypes and Demographic History

  • Ferreri, Miro;Gao, Jian;Wang, Zhi;Chen, Liben;Su, Jingliang;Han, Bo
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.24 no.8
    • /
    • pp.1048-1052
    • /
    • 2011
  • The Chinese Holstein cattle breed, an introduced breed in China, has been crossbred with native cattle breeds. We hypothesised that the Chinese Holstein local population in Beijing share haplotypes with native Asian cattle breeds, the result of a sudden population expansion in the recent past. We also hypothesised that crossbreeding and population expansion left traces that shaped the genetic makeup of the breed. Evaluation of this was performed by mitochondrial DNA (mtDNA) sequence analysis of Chinese Holstein cattle from Beijing (n = 41) and a comparison of them with the published mtDNA sequences (n = 293) of 14 Asian breeds with an emphasis on Chinese native cattle breeds. Three shared common haplotypes between Chinese Holstein cattle and native Asian cattle were found. Moreover, a high level of haplotype diversity in Chinese Holstein cattle (h = 0.9557) and low nucleotide diversity (${\pi}$ = 0.0052) was found, indicating a past population bottleneck followed by rapid population growth. These findings are supported by the significantly negative deviation of Tajima's D (-1.82085), the star-like pattern of dominant haplotypes and the pairwise mismatch distribution analysis, which showed a unimodal pattern.

Increase in Linolenate Contents by Expression of the fad3 Gene in Transgenic Tobacco Plants

  • Kang, Young-Hwi;Min, Bok-Kee;Park, Hee-Sung;Lim, Kyung-Jun;Huh, Tae-Lin;Lee, Se-Yong
    • BMB Reports
    • /
    • v.29 no.4
    • /
    • pp.308-313
    • /
    • 1996
  • An 1.4 kb of the fad3 cDNA encoding microsomal linoleic acid desaturase catalyzing the conversion of linoleic acid (18:2, ${\omega}-6$) to linolenic acid (18:2, ${\omega}-3$) was introduced into tobacco plants by the Agrobacterium-mediated plant transformation, Among the transgenic tobacco plants conferring kanamycin resistance, five transformants showing increment in unsaturated fatty acid contents were selected and further analyzed for the transgenecity, In genomic Southern blot analyses, copy numbers of the integrated fad3 DNA in chromosomal DNA of the five transgenic tobacco plants were varied among the transgenic lines. By Northern blot analyses, the abundancy of the fad3 mRNA transcript directed by Cauliflower Mosaic Virus 35S promoter was consistent with the relative copy number of the fad3 DNA integrated in the chromosome of transgenic tobacco plants. When compared with the wild type, accumulation of linolenic acid in transgenic tobacco roots was elevated 3.7- to 4.7-fold showing a corresponding decrease in the linoleic acid contents; however, slight increments for linolenic acid were noticed in transgenic leaf tissues. These results indicated that the elevated level of fad3 expression is achieved in transgenic tobacco plants.

  • PDF

Mammary Gland-Specific Expression of Biologically Active Human Osteoprotegerin in Transgenic Mice

  • Sung, Yoon-Young;Lee, Chul-Sang
    • Development and Reproduction
    • /
    • v.17 no.1
    • /
    • pp.1-8
    • /
    • 2013
  • Osteoprotegerin (OPG) is a secreted glycoprotein that regulates bone resorption by inhibiting differentiation and activation of osteoclast, thereby potentially useful for the treatment of many bone diseases associated with increased bone loss. In this study, we designed a novel cDNA expression cassette by modifying the potent and mammary gland-specific goat ${\beta}$-casein/hGH hybrid gene construct and examined human OPG (hOPG) cDNA expression in transgenic mice. Six transgenic mice all successfully expressed hOPG in their milk at the level of 0.06-2,000 ${\mu}g/ml$. An estimated molecular weight of the milk hOPG was 55 kDa in SDS-PAGE, which is the same as a naturally glycosylated monomer. This hOPG expression was highly specific to the mammary glands of transgenic mice. hOPG mRNA was not detected in any organs analyzed except mammary gland. Functional integrity of milk hOPG was evaluated by TRAP (tartrate-resistant acid phosphatase) activity assay in bone marrow cell cultures. OPG ligand (OPG-L) treatment increased TRAP activity by two fold but it was completely abolished by co-treatment with transgenic milk containing hOPG. Taken together, our novel cDNA expression cassette could direct an efficient expression of biologically active hOPG, a potential candidate pharmaceutical for bone diseases, only in the mammary gland of transgenic mice.

Complementary DNA Cloning and Sequencing of the Coat Protein Gene of Potato Virus Y-Ordinary Korean Strain (감자바이러스 Y의 OK계통에 대한 외피단백질 유전자 cDNA 클로닝 및 염기서열 분석)

  • 정승룡;최장경;길전행이;이부영
    • Korean Journal Plant Pathology
    • /
    • v.11 no.1
    • /
    • pp.73-79
    • /
    • 1995
  • Complementary DNAs (cDNAs) to the coat protein gene of an ordinary Korean strain of potato virus Y (PVY-OK) isolated from potato (cv. Superior) were synthesized and cloned into a plasmid pUC119 and sequenced. The RNA of the virus propagated in tobacco (Nicotinaa sylvestris) was extracted by the method of phenol extraction. The first strand of cDNAs to the coat protein penomic RNA of the virus was made by Moloney murine leukemia virus reverse transcriptase. The cDNA were synthesized and amplified by the method of polymerase chain reaction (PCR) using a pair of oligonucleotide primers. PVYCP3P and PVYCP3M. The size of cDNAs inserted in pUC119 plasmid was estimated as about 840 bp upon agarose gel electrophoresis. Double stranded cDNAs were transformed into the competent cell of E. coli JM109. Sequence analysis of cDNAs was conducted by the dideoxynucleotide chain termination method. Homology of cDNAs of the PVY-OK coat protein genomic RNA with those of PVY-O (Japan), PVY-T (Japan), PVY-TH (Japan), PVYN (The Netherlands),and PVYY (France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. Homology at the amino acid level turned out to the be 97.4%, 92.5%, 92.9%, 92.9% and 98.5%, respectively.

  • PDF

Isolation and Characterization of a Cdna ( Fp 1 ) Encoding the Iron Storage Protein in Red Pepper ( Capsicum annuum L. )

  • Kim, Ho-Young;Lee, Young-Ok;Noh, Ill-Sup;Kang, Hee-Wan;Kameya, Toshiaki;Saito, Takashi;Kang, Kwon-Kyoo
    • Plant Resources
    • /
    • v.1 no.1
    • /
    • pp.13-21
    • /
    • 1998
  • A cDNA Fragment encoding iron storage protrin generated by polymerase chain reaction(PCR) using highly conserved regions of ferritin related genes were used to sereen a red pepper cDNA library. cDNA clone was designated as Fp1. Fp1 clone contatines a 5' nontranslated region of 51dp containing stop conds. Down stream from 5' UTP. an open reading frame of 750bp was observed. followed by a 3' UTR of 272bp. The deduces amino acid sequence of red pepper protein(Fp1) showed 84%, 48% and 36% identity with soybean(SolC). human(HuL H) and horse spleen(HoS-L) ferritin mRNA accumulation in response to iron. Ferritin mRNA accumulation was transient and particularly abundant in leaves. reaching a maxmum at 12h. The level of ferritin mRNA in roots was affected to a lesser extent than in leaves.

  • PDF

Gene Expression study of human chromosomal aneuploid

  • Lee Su-Man
    • Proceedings of the Korean Society for Bioinformatics Conference
    • /
    • 2006.02a
    • /
    • pp.98-107
    • /
    • 2006
  • Chromosomal copy number changes (aneuploidies) are common in human populations. The extra chromosome can affect gene expression by whole-genome level. By gene expression microarray analysis, we want to find aberrant gene expression due to aneuploidies in Klinefelter (+X) and Down syndrome (+21). We have analyzed the inactivation status of X-linked genes in Klinefelter Syndrome (KS) by using X-linked cDNA microarray and cSNP analysis. We analyzed the expression of 190 X-linked genes by cDNA microarray from the lymphocytes of five KS patients and five females (XX) with normal males (XY) controls. cDNA microarray experiments and cSNP analysis showed the differentially expressed genes were similar between KS and XX cases. To analyze the differential gene expressions in Down Syndrome (DS), Amniotic Fluid (AF)cells were collected from 12 pregnancies at $16{\sim}18$ weeks of gestation in DS (n=6) and normal (n=6) subjects. We also analysis AF cells for a DNA microarray system and compared the chip data with two dimensional protein gel analysis of amniotic fluid. Our data may provide the basis for a more systematic identification of biological markers of fetal DS, thus leading to an improved understanding of pathogenesis for fetal DS.

  • PDF

Cloning and Nucleotide Sequence of a cDNA Encoding the Rat Triosephosphate Isomerase

  • Lee, Kyunglim;Ryu, Jiwon
    • Archives of Pharmacal Research
    • /
    • v.19 no.6
    • /
    • pp.497-501
    • /
    • 1996
  • A gene coding for triosephosphate isomerase (TPI) from a rat skeletal muscle cDNA library was cloned and its nucleotide sequence was determined. The 1, 348-bp cDNA clone contains 24 bp $5^I$ noncoding region, the entire 750 bp coding region corresponding to a protein of 249 amino acids, $547bp 3^I$ noncoding region and part of a poly(A) tail. It also contains a polyadenylation signal, AATAAA, starting from 17 bp upstream of the poly(A) tail. The calculated molecular weight of rat TPI is 27.8 kDa and the net charge is +4. The deduced amino acid sequence from rat TPI CDNA sequence has 93% and 94% homology with that of mouse and human clones, respectively. The amino acids at the residue of Asn12, Lys14, His96, Glu 166, His96, His101, Ala177, Tyr165, Glu13O, Tyr2O9, and Ser212 in catalytic site are completely identical, confirming that the functional residues in TPI proteins are highly conserved throughout evolution. The most profound characteristic of rat TPI enzyme, compared with other TPIs, is that there are five cysteine substitutions at the residue of 21, 27, 159, 195 and 204. A Glu123 instead of Gly was found in rabbit, rhesus, mouse and human sequences. Through the method of RT-PCR, the mRNA transcription level of TPI gene was found to be different among various tissues and was highest in muscle.

  • PDF