• Journal of Life Science
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• v.26 no.4
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• pp.419-430
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• 2016

Powdery mildew disease caused by Podosphaera xanthii is a major concern for Cucumis melo production worldwide. Knowledge on genetic behavior of the related genes and their modulating phytohormones often offer the most efficient approach to develop resistance against different diseases. Mildew Resistance Locus O (MLO) genes encode proteins with seven transmembrane domains that have significant function in plant resistance to powdery mildew fungus. We collected 14 MLO genes from ‘Melonomics’ database. Multiple sequence analysis of MLO proteins revealed the existence of both evolutionary conserved cysteine and proline residues. Moreover, natural genetic variation in conserved amino acids and their replacement by other amino acids are also observed. Real-time quantitative PCR expression analysis was conducted for the leaf samples of P. xanthii infected and phyto-hormones (methyl jasmonate and salicylic acid) treated plants in melon ‘SCNU1154’ line. Upon P. xanthii infection using 7 different races, the melon line showed variable disease reactions with respect to spread of infection symptoms and disease severity. Three out of 14 CmMLO genes were up-regulated and 7 were down-regulated in leaf samples in response to all races. The up- or down-regulation of the other 4 CmMLO genes was race-specific. The expression of 14 CmMLO genes under methyl jasmonate and salicylic acid application was also variable. Eleven CmMLO genes were up-regulated under salicylic acid treatment, and 7 were up-regulated under methyl jasmonate treatments in C. melo L. Taken together, these stress-responsive CmMLO genes might be useful resources for the development of powdery mildew disease resistant C. melo L.

• Horticultural Science & Technology
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• v.34 no.2
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• pp.324-330
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• 2016

To determine whether FT-IR spectral analysis based on multivariate analysis for whole cell extracts can be used to discriminate between artichoke (Cynara cardunculus var. scolymus L.) plants at the metabolic level, leaves of ten artichoke plants were subjected to Fourier transform infrared(FT-IR) spectroscopy. FT-IR spectral data from leaves were analyzed by principal component analysis (PCA), partial least square discriminant analysis (PLS-DA) and hierarchical clustering analysis (HCA). FT-IR spectra confirmed typical spectral differences between the frequency regions of 1,700-1,500, 1,500-1,300 and $1,100-950cm^{-1}$, respectively. These spectral regions reflect the quantitative and qualitative variations of amide I, II from amino acids and proteins ($1,700-1,500cm^{-1}$), phosphodiester groups from nucleic acid and phospholipid ($1,500-1,300cm^{-1}$) and carbohydrate compounds ($1,100-950cm^{-1}$). PCA revealed separate clusters that corresponded to their species relationship. Thus, PCA could be used to distinguish between artichoke species with different metabolite contents. PLS-DA showed similar species classification of artichoke. Furthermore these metabolic discrimination systems could be used for the rapid selection and classification of useful artichoke cultivars.

• BMB Reports
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• v.45 no.3
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• pp.183-188
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• 2012

Participates in actin remodeling through Rac and receptor endocytosis via Rab5. Here, we used yeast two-hybrid system with Eps8 as bait to screen a human brain cDNA library. ITSN2 was identified as the novel binding factor of Eps8. The interaction between ITSN2 and Eps8 was demonstrated by the in vivo co-immunoprecipitation and colocalization assays and the in vitro GST pull-down assays. Furthermore, we mapped the interaction domains to the region between amino acids 260-306 of Eps8 and the coiled-coil domain of ITSN2. In addition, protein stability assays and immunofluorescence analysis showed ITSN2 overexpression induced the degradation of Eps8 proteins, which was markedly alleviated with the lysosome inhibitor NH4Cl treatment. Taken together, our results suggested ITSN2 interacts with Eps8 and stimulates the degradation of Eps8 proteins.

• Journal of Animal Science and Technology
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• v.46 no.6
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• pp.909-916
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• 2004

CCAAT/enhancer binding proteins(C/EBP) are a group of transcription factors expressed during preadipocyte differentiation. In the C/EBPs, C/EBPa plays an important role in lipid deposition and adipocyte differentiation. In this studies, we report the identification, characterization, and expression of a Hanwoo CIEBP$\alpha$ The Hanwoo C/EBP$\alpha$DNA includes a 1059 bp open reading frame encoding a protein of 353 amino acids. The CIEBPa amino acid sequences of the Hanwoo show strong conservation with the corresponding sequences reported in other species. The distribution of C/EBP$\alpha$ mRNA in various tissues of Hanwoo aged 12 months were investigated using Northern blotting analysis. The highest expression was detected in adipose tissue and more lower expression was detected in colon and lung. We also identified expression of C/EBPa mRNA in Hanwoo sirloin and adipose tissue aged 12, 26, and 30 months by real-time RT-PCR. The higest expression were detected at 26 months in the sirloin and at 12 and 26 months in the adipose tissue.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.23
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• pp.10255-10262
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• 2015

High risk human papillomavirus (HR-HPV) E2 proteins play roles in transcriptional regulation and are commonly functionally disrupted when the HPV genome integrates into host chromosomes. Some 15-40% of cancer cases, however, contain an intact E2 gene or episomal HPV. In these cases, polymorphism of the E2 gene might be involved. This study aimed to determine polymorphisms of the E2 gene in episomal HPV16 detected in high grade squamous intraepithelial lesions and squamous cell carcinomas and altered functions compared to the E2 prototype. The E2 gene was amplified and sequenced. Two expression vectors containing E2 gene polymorphisms were constructed and transfected in SiHa and C33A cells, then E6 gene as well as Il-10 and TNF-${\alpha}$ expression was determined by quantitative RT-PCR. Expression vectors and reporter vectors containing the HPV16 long control region (LCR) were co-transfected and transcriptional activity was determined. The results showed that a total of 32 nucleotides and 23 amino acids were changed in all 20 cases of study, found in the transactivation (TA) domain, hinge (H) region and DNA binding (DB) domain with 14, 5 and 13 nucleotide positions. They mostly caused amino acid change. The expressing vectors containing different E2 gene polymorphisms showed E6 mRNA suppression, TNF-${\alpha}$ mRNA suppression and IL-10 induction but no statistically significant differences when compared to the E2 prototype. Moreover, promoter activity in HPV16 LCR was not affected by E2 protein with different gene polymorphisms, in contrast to nucleotide variations in LCR that showed an effect on transcription activity. These results demonstrated that E2 gene polymorphisms of episomal HPV16 did not affect transcriptional regulation and suggested that nucleotide variation as well as epigenetic modification of the LCR might play a role in inducing malignant transformation of cells containing episomal HPV16.

• Journal of Microbiology
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• v.45 no.1
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• pp.21-28
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• 2007

In order to search for specific genotypes related to this unique phenotype, we used whole genomic DNA microarray to characterize the genomic diversity of Helicobacter pylori (H. pylori) strains isolated from clinical patients in China. The open reading frame (ORF) fragments on our microarray were generated by PCR using gene-specific primers. Genomic DNA of H. pylori 26695 and J99 were used as templates. Thirty-four H. pylori isolates were obtained from patients in Shanghai. Results were judged based on In(x) transformed and normalized Cy3/Cy5 ratios. Our microarray included 1882 DNA fragments corresponding to 1636 ORFs of both sequenced H. pylori strains. Cluster analysis, revealed two diverse regions in the H. pylori genome that were not present in other isolates. Among the 1636 genes, 1091 (66.7%) were common to all H. pylori strains, representing the functional core of the genome. Most of the genes found in the H. pylori functional core were responsible for metabolism, cellular processes, transcription and biosynthesis of amino acids, functions that are essential to H. pylori's growth and colonization in its host. In contrast, 522 (31.9%) genes were strain-specific genes that were missing from at least one strain of H. pylori. Strain-specific genes primarily included restriction modification system components, transposase genes, hypothetical proteins and outer membrane proteins. These strain-specific genes may aid the bacteria under specific circumstances during their long-term infection in genetically diverse hosts. Our results suggest 34 H. pylori clinical strains have extensive genomic diversity. Core genes and strain-specific genes both play essential roles in H. pylori propagation and pathogenesis. Our microarray experiment may help select relatively significant genes for further research on the pathogenicity of H. pylori and development of a vaccine for H. pylori.

• Reproductive and Developmental Biology
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• v.34 no.1
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• pp.33-40
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• 2010

Equine chorionic gonadotropin (eCG) is a heavily glycosylated glycoprotein composed of non-covalently linked $\alpha$- and $\beta$-subunits. To study the function and signal transduction of tethered recombinant-eCG (rec-eCG), a single chain eCG molecule was constructed, and the rec-eCG protein was prepared. In this study, we constructed 5 mutants (${\Delta}1$, ${\Delta}2$, ${\Delta}3$, ${\Delta}4$, and ${\Delta}5$) of rec-eCG using data about known glycoprotein hormones to analyze the role of specific follicle stimulating homone (FSH)-like activity. Three amino acids of certain specific sites were replaced with alanine. The expression vectors were transfected into CHO cells and subjected to G418 selection for 2~3 weeks. The media were collected and the quantity of secreted tethered rec-eCGs was quantified by ELISA. The LH- and FSH-like activities were assayed in terms of cAMP production by rat LH/CG and rat FSH receptors. Then, the metabolic clearance rate analyzed by the injection of rec-eCG (5 IU) into the tail vein was analyzed. The mutant eCGs (${\Delta}l$, ${\Delta}4$, and ${\Delta}5$) were transcripted, but not translated into proteins. Rec-eCG A2 was secreted in much lower amounts than the wild type. Only the rec-eCG ${\Delta}3$ ($\beta$-subunit: $Gln^{94}-Ile^{95}-Lys^{96}{\rightarrow}Ala^{94}-Ala^{95}-Ala^{96}$) was efficiently secreted. Although activity is low, its LH-like activity was similar to that of tethered $eCG{\beta\alpha}$. However, the FSH-like activity of rec-$eCG{\beta\alpha\Delta}3$ was completely flat. The result of the analysis of the metabolic clearance rate shoed the persistence of the mutant in the blood until 4 hours after the injection. After then, it almost disappeared at 8 hours. Taken together, these data suggest that 94~96 amino acid sequences in eCG $\beta$-subunit appear to be of utmost importance for signal transduction of the FSH receptor.

• Korean Journal of Food Science and Technology
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• v.30 no.6
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• pp.1259-1266
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• 1998

To clarify the values and varieties of the buckwheats as a dietary source of nutritional and functional components, thirteen different samples of buckwheat were analyzed for this investigation. Six developed seeds were given by RDA, Korea or RDA branch of Kangwondo, and seven land race seeds were collected from a farmhouse. Amino acid analysis showed that glutamate, arginine and asparagine were major amino acids, whereas tryptophan, methionine and cysteine were minor ones of buckwheat. In addition, tryptophan content of buckwheat cultivars from Korea was 195 mg% on average. The content of rutin tended to be higher in developed cultivars than land races. On the other hand, the contents of phytic acid in buckwheats were in the range of 7.0 to 13.6 mg/g. In the tocopherol homologues of the buckwheats analyzed by HPLC, mean ${\gamma}-tocopherol$ contents were 6.16 mg/100 g with the actual range of $4.67{\sim}8.58\;mg/100g$, whereas ${\beta}-form$ was very low or zero. There were a big variations in the iron content of the buckwheats of the minerals. SDS-PAGE showed that total proteins from buckwheats exhibited a relatively similar electrophoretic patterns on the whole. The results show that CV Suwon 1 has good quality, judged from the distribution of the components of buckwheats analyzed.

• Korean Journal of Food Science and Technology
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• v.14 no.3
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• pp.250-256
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• 1982

A series of studies were conducted to find out the possibility of utilizing grape seed as resources of food fats and proteins, and the results of the studies are as follows: The grape seed contained 25.1%, of crude fat and 12.0% of crude protein. The lipid, fractions obtained by silicic acid column chromatography were mainly composed of about 95.5% neutral lipid, whereas compound lipid was only 4.5% level. Among the neutral lipid by thin layer chromatography, triglyceride was 91.89%, sterol ester, sterol, diglyceride and free fatty acid were 3.24%, 2.87%, 1.20% and 0.80%, respectively The predominant fatty acids of total and neutral lipids were linoleic acid $(69.72{\sim}71.72%)$ and oleic acid $18.09{\sim}19.46%)$, but those of glycolipid and phospolipid were linoleic acid $(31.49{\sim}38.18%)$, oleic acid $(20.20{\sim}35.27%)$ and palmitic acid $(26.80{\sim}39.98%)$. The major fatty acids of triglyceride separated from neutral lipid were oleic acid (43.08%), linoleic acid (38.42%) and palmitic acid (11.60%). The salt soluble protein of grape seed was highly dispersible in 0.02M sodium phosphate buffer containing about 1.0M $MgSO_4$, and the extractability of seed protein was 31%. Glutamic acid was the major amino acid in salt soluble protein, followed by arginine and aspartic acid. The electrophoretic analysis showed 3 bands in grape seed protein, and the collection rate of the main protein fraction purified by Sephadex G-100 and G-200 was 82%. Glutamic acid, aspartic acid and arginine were the major amino acids of the main grape seed protein. The molecular weight for the main protein of the grape seed was estimated to be 81,000.

• Journal of the Korean Society of Food Science and Nutrition
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• v.12 no.1
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• pp.46-50
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• 1983

In order to find out the possibility of utilizing red pepper seed as food resources of fats and proteins, a series of studies were conducted. The red pepper seed contained 27.6% of crude fat and 22.2% of crude protein. The lipid fractions obtained by silicic acid column chromatography were mainly composed of 95.4% neutral lipid, where as compound lipid were 4.6%. Among the neutral lipid separated by thin layer chromatography, triglyceride was 85.6%, sterol ester 4.9%, free fatty acids 3.4%, diglyceride 2.5%, sterol 2.2% and monoglyceride 1.1%, respectively. The predominant fatty acids of red pepper seed oil were linoleic acid (57.1-75.4%), palmitic acid (13.9-21.3%) and oleic acid (8.0-15.1%), especially glycolipid contained 1.7% of linolenic acid and small amount of myristic acid and arachidic acid. The salt soluble protein of red pepper seed was highly dispersible in 0.02M sodium phosphate buffer containing 1.0M $MgSO_4$, and the extractability of seed protein was about 25.0%. Glutamic acid and arginine were major amino acids of red pepper seed protein. The electrophoretic analysis showed 6 bands in seed protein, and the collection rate of the main protein fraction purified by sephadex G-100 and G-200 was about 62.2%. Glutamic acid (19.9%) was major amino acid of the main protein, followed by glycine and alanine. The molecular weight of the main protein was estimated to be 93,000.