• Title/Summary/Keyword: F gene

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Characterization of the xaiF Gene Encoding a Novel Xylanase-activity- increasing Factor, XaiF

  • Cho, Ssang-Goo;Choi, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.8 no.4
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    • pp.378-387
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    • 1998
  • The DNA sequence immediately following the xynA gene of Bacillus stearothermophilus 236 [about l-kb region downstream from the translational termination codon (TAA) of the xynA gene]was found to have an ability to enhance the xylanase activity of the upstream xynA gene. An 849-bp ORF was identified in the downstream region, and the ORF was confirmed to encode a novel protein of 283 amino acids designated as XaiF (xylanase-activity-increasing factor). From the nucleotide sequence of the xaiF gene, the molecular mass and pI of XaiF were deduced to be 32,006 Da and 4.46, respectively. XaiF was overproduced in the E. coli cells from the cloned xaiF gene by using the T7 expression system. The transcriptional initiation site was determined by primer extension analysis and the putative promoter and ribosome binding regions were also identified. Blast search showed that the xaiF and its protein product had no homology with any gene nor any protein reported so far. Also, in B. subtilis, the xaiF trans-activated the xylanase activity at the same rate as in E. coli. In contrast, xaiF had no activating effect on the co-expressed ${\beta}-xylosidase$ of the xylA gene derived from the same strain of B. stearothermophilus. In addition, the intracellular and extracellular fractions from the E. coli cells carrying the plasmid-borne xaiF gene did not increase the isolated xylanase activity, indicating that the protein-protein interaction between XynA and XaiF was not a causative event for the xylanase activating effect of the xaiF gene.

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Effects of TESTIN Gene Expression on Proliferation and Migration of the 5-8F Nasopharyngeal Carcinoma Cell Line

  • Zhong, Zhun;Zhang, Fei;Yin, Shu-Cheng
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.6
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    • pp.2555-2559
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    • 2015
  • Purpose: To investigate effects of the TESTIN (TES) gene on proliferation and migration of highly metastatic nasopharyngeal carcinoma cell line 5-8F and the related mechanisms. Materials and Methods: The target gene of human nasopharyngeal carcinoma cell line 5-8F was amplified by PCR and cloned into the empty plasmid pEGFP-N1 to construct a eukaryotic expression vector pEGFP-N1-TES. This was then transfected into 5-8F cells. MTT assays, flow cytometry and scratch wound tests were used to detect the proliferation and migration of transfected 5-8F cells. Results: A cell model with stable and high expression of TES gene was successfully established. MTT assays showed that the OD value of 5-8F/TES cells was markedly lower than that of 5-8F/GFP cells and 5-8F cells (p<0.05). Flow cytometry showed that the apoptosis rate of 5-8F/TES cells was prominently increased compared with 5-8F/GFP cells and 5-8F cells (p<0.05). In vitro scratch wound assays showed that, the width of the wound area of 5-8F/TES cells narrowed slightly, while the width of the wound area of 5-8F/ GFP cells and 5-8F cells narrowed sharply, suggesting that the TES overexpression could inhibit the migration ability. Conclusions: TES gene expression remarkably inhibits the proliferation of human nasopharyngeal carcinoma cell line 5-8F and reduces its migration in vitro. Thus, it may be a potential tumor suppressor gene for nasopharyngeal carcinoma.

Inheritance of Mammoth Gene and White Flower in Flue-cured Tobacco(Nicotiana tabacum L.) (황색종 연초에서 mammoth gene과 흰꽃의 유전)

  • 조수헌
    • Journal of the Korean Society of Tobacco Science
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    • v.22 no.1
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    • pp.71-75
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    • 2000
  • The genetic makeup could be the most important among many factors affecting yield and quality of tobacco(Nicotiana tabacum L.). The mammoth gene found in N. tabacum is associated with greater leaf number and poor leaf quality. This study was carried out to obtain the basic information about the inheritance of mammoth gene and white flower color. Two flue-cured breeding lines, KF 9373-2 and KF 8832-85, F$_1$, F$_2$, two parents backcrossed with F$_1$, and F$_3$ lines derived from cross of above two lines were investigated for flowering type(mammoth gene) and flower color. All plants of F$_1$ population revealed normal flowering type and pink flower color. The progeny of F$_2$ generation was segregated into the phenotypic ratio of 9 : 3 : 3 : 1 with normal flowering type and pink flower color, normal and white, non flowering type(NF) and pink, and NF and white, respectively. Among the progenies of back-crossing populations, the flowering type showed a segregation ratio of 1 : 1 as normal and NF in BP$_1$ and flower color did also 1 : 1 as pink and white in BP$_2$. All lines have the mammoth gene in F$_3$. that were selected in F$_2$ progeny as non flowering. But 9 lines among 14 were segregated with 3 : 1 as pink and white flower in F$_3$. which were selected in F$_2$ as pink flower color. These results indicated that the characters of mammoth gene and white flower were controlled by a pair of recessive genes, respectively.

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Genome of Betaproteobacterium Caenimonas sp. Strain SL110 Contains a Coenzyme $F_{420}$ Biosynthesis Gene Cluster

  • Li, Xiuling;Feng, Fuying;Zeng, Yonghui
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1490-1494
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    • 2014
  • To probe the genomic properties of microbes thriving in desert lakes, we sequenced the full genome of a betaproteobacterial strain (SL110) belonging to the understudied genus Caenimonas of the family Comamonadaceae. This strain was isolated from a freshwater lake in the western Gobi Desert, Northern China. Its genome contains genes encoding carbon monoxide dehydrogenase, nitrate reductase, nitrite reductase, nitric oxide reductase, and sulfur oxidation enzymes, highlighting the potentially important contribution of this group of bacteria to the cycling of inorganic elements in nature. Unexpectedly, a coenzyme $F_{420}$ biosynthesis gene cluster was identified. A further search for $F_{420}$ biosynthesis gene homologs in genomic databases suggests the possible widespread presence of $F_{420}$ biosynthesis gene clusters in proteobacterial genomes.

Skin Improvement Effect of Mask Pack with Snail Fermented Extract (달팽이 발효 추출물을 이용한 마스크 팩의 피부개선효과)

  • Zhoh, Choon-Koo;Lee, Min-Hee;Kim, In-Young;Shin, Ji-Young
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.1
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    • pp.30-40
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    • 2016
  • This study related to get skin improvement effect of sheet mask pack using 5% of $2^{nd}$ fermented snail extract. To the scientific study of the skin, it was used for this sheet mask of 100% cotton as cutting face shape. We reported the results of measurement of skin improvement effect with the placebo and Gene-SEM (general snail extract mask). First; moisturizing effect of 2F-SEM ($2^{nd}$ fermented snail extract mask) has been increased about 11% than the PM and Gene-SEM was improved about 4.7% more than the PM (placebo mask). Second; elasticity of the mask pack containing $2^{nd}$ fermented snail extract was 13.8% better than PM and Gene-SEM was improved about 6.7 % more than PM. Third; Skin roughness of 2F-SEM has improved 6.80% than PM and Gene-SEM was improved about 2.3% more than PM. Fourth; melanin reductive effect of 2F-SEM was improved about15.0% more than PM and Gene-SEM was improved about 8.7% more than PM. Fifth; fine wrinkle decline effect of 2F-SEM was about 8.0% better than PM and Gene-SEM was improved with 5.1% more than PM. Sixth; sensorial evaluation of 2F-SEM using $2^{nd}$ fermented snail extract was significantly showed difference merits regarding softness, moisture, fine-wrinkle improvement.

Studies on QTLs for Bakanae Disease Resistance with Populations Derived from Crosses between Korean japonica Rice Varieties

  • Dong-Kyung Yoon;Chaewon Lee;Kyeong-Seong Cheon;Yunji Shin;Hyoja Oh;Jeongho Baek;Song-Lim Kim;Young-Soon Cha;Kyung-Hwan Kim;Hyeonso Ji
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.201-201
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    • 2022
  • Rice bakanae disease is a serious global threat in major rice-cultivating regions worldwide causing high yield loss. It is caused by the fungal pathogen Fusarium fujikuroi. Varying degree of resistance or susceptibility to bakanae disease had been reported among Korean japonica rice varieties. We developed a modified in vitro bakanae disease bioassay method and tested 31 Korean japonica rice varieties. Nampyeong and Samgwang varieties showed highest resistance while 14 varieties including Junam and Hopum were highly susceptible with 100% mortality rate. We carried out mapping QTLs for bakanae disease resistance with four F2:F3 populations derived from the crosses between Korean japonica rice varieties. The Kompetitive Allele-Specific PCR (KASP) markers developed in our laboratory based on the SNPs detected in Korean japonica rice varieties were used in genotyping F2 plants in the populations. We found four major QTLs on chromosome 1, 4, 6, and 9 with LOD scores of 21.4, 6.9, 6.0, and 60.3, respectively. In addition, we are doing map-based cloning of the QTLs on chromosome 1 and 9 which were found with Junam/Nampyeong F2:F3 population and Junam/Samgwang F2:F3 population, respectively. These QTLs will be very useful in developing bakanae disease resistant high quality rice varieties.

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Segregation Mode of Plant Height in Crosses of Rice Cultivars Ⅸ. Crosses between Semi-dwarf Japonicas and Semi-dwarf(d-t) gene Testers (수도 품종간 교잡에 있어서 간장의 유전분리 Ⅸ. 단간 Japonica 품종과 Semi-dwarf (d-t) gene 검정친과의 조합)

  • Kim, Yong-Kwon;Kim, Hong-Yeol;Nam, Yeong-Woo;Park, Sun-Zik;Heu, Mun-Hue
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.30 no.4
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    • pp.449-454
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    • 1985
  • In order to search for the semi-dwarf japonica varieties allelic to the semi-dwarf rice cultivar which is controlled by d-t gene, seven dwarf japonica varieties. Reimei, Hoyoku. Shiranui, Kokumasari, M 7. S.224 and S.295 were crossed to the semi-dwarf cultivar, wx 817. wx 817 is known to have semi-dwarf gene d-t. Their F$_1$, F$_2$ and F$_3$ were grown in 1984 and 1985 and culm lengths were measured at harvest. The results are summarized as follows. 1. The F$_2$s of all 7 cross combinations showed normal distribution and no segregation. 2. The range of culm length variation in the F$_3$ was variable depending on the cross combination, but the general pattern was similar in the all 7 crosses. 3. The mean of F$_3$ and parental F$_2$ mean which were selected into short, medium and tall groups were similar and showed no segregation, implying the selection efficiency in F$_2$. 4. From the results of F$_2$ and F$_3$ segregations, it is concluded that the culm length of the 7 semi-dwarf japonicas tested here are controlled by the same major gene d-t although they are modified by different minor genes.

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Characteristics of Oncolytic Adenovirus Replication and Gene Expression in Hypoxic Condition

  • Kim, Hong-Sung
    • Biomedical Science Letters
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    • v.17 no.3
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    • pp.185-190
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    • 2011
  • Adenovirus type 5 (Ad5) vectors have been used for gene transfer to a wide variety of cell types in vivo and in vitro. The advantages of adenovirus vectors include the high titer of virus readily obtained in large scale preparations, their ability to transduce dividing and non dividing cells, and the high level of transgene expression. Since adenovirus vectors do not integrate in host cell DNA, there is a lack of insertional mutagenesis. However, many human tumor cells lack expression of the adenovirus 5 receptors and contain areas of hypoxia. In order to identify the pattern of replication and gene expression of oncolytic adenovirus in hypoxic condition, multiple different fiber modified Ads (Ad5F/S11, Ad5F/S35, Ad5F/K7, Ad5F/K21, and Ad5F/RGD) was compared. The replication of all fiber modified adenovirus was inhibited in hypoxic condition in HEK 293 cells, but gene expression has variety on different tumor cell lines and the level of coxackievirus and adenovirus receptor (CAR) expression. These data suggest that CAR expression pattern and hypoxic condition of tumor are considered for optimal oncolytic adenovirus application.

Molecular Cloning, Sequencing, and Expression of a Fibrinolytic Serine-protease Gene from the Earthworm Lumbricus rubellus

  • Cho, Il-Hwan;Choi, Eui-Sung;Lee, Hyung-Hoan
    • BMB Reports
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    • v.37 no.5
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    • pp.574-581
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    • 2004
  • The full-length cDNA of the lumbrokinase fraction 6 (F6) protease gene of Lumbricus rubellus was amplified using an mRNA template, sequenced and expressed in E. coli cells. The F6 protease gene consisted of pro- and mature sequences by gene sequence analysis, and the protease was translated and modified into active mature polypeptide by N-terminal amino acid sequence analysis of the F6 protease. The pro-region of F6 protease consisted of the 44 residues from methionine-1 to lysine-44, and the mature polypeptide sequence (239 amino acid residues and one stop codon; 720 bp) started from isoleucine-45 and continued to the terminal residue. F6 protease gene clones having pro-mature sequence and mature sequence produced inclusion bodies in E. coli cells. When inclusion bodies were orally administrated rats, generated thrombus weight in the rat' venous was reduced by approximately 60% versus controls. When the inclusion bodies were solubilized in pepsin and/or trypsin solutions, the solubilized enzymes showed hemolytic activity in vitro. It was concluded the F6 protease has hemolytic activity, and that it is composed of pro- and mature regions.

Effect of Lipid Compositions on Gene Transfer into 293 Cells Using Sendai F/HN-virosomes

  • Kim, Hong-Sung;Park, Yong-Serk
    • BMB Reports
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    • v.35 no.5
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    • pp.459-464
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    • 2002
  • Fusogenic liposomes that incorporate Sendai virus envelope proteins, so-called Sendai virosomes, have been developed for in vitro and in vivo genetic modification of animal cells. In this study, several different virosomes of varying lipid compositions were formulated and their in vitro gene-transfer efficiencies compared. The virosomes were prepared by quantitative reconstitution of the Sendai envelope, fusion (F) and hemagglutinin-neuraminidase (HN) proteins into liposomal vesicles. Virosomes that contained luciferase reporter genes were tested in 293 transformed human kidney cells. F/HN-virosomes that were prepared with an artificial Sendai viral envelope (ASVE-virosomes) or phosphatidylserine (PS-virosomes) exhibited an 8- or 6-fold higher gene-transfer efficiency than cationic liposomes that were made with 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP). F/HN-virosomes that were prepared with phosphatidic acid (PA-virosomes) instead of PS were less efficient in gene transfer than either ASVE- or PS-virosomes. In addition, the genetransfer capability of ASVE- and PS-virosomes was maximal at a $Ca^{2+}$ concentration of 510 mM. These results suggest that the incorporated lipid components significantly affect the in vitro gene transfer that is mediated by Sendai F/HN-virosomes.