• 대한의생명과학회지
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• 제19권3호
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• pp.239-244
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• 2013

Serum liver enzyme levels are widely used in the clinical diagnosis of liver diseases and the assessment of liver status. They also have epidemiological significance to be prospective risk factors for type 2 diabetes, cardiovascular disease. In the previous study, single nucleotide polymorphisms (SNPs) in several genes have been reported to be associated with serum liver enzyme levels in American population. We aimed to confirm whether the genetic variation of C16orf47 (chromosome 16 open reading frame 47) gene also influence the serum liver enzyme levels in Korean population. We genotyped variants in or near C16orf47 in a population-based sample including 994 unrelated Korean adult. Here, we performed association analysis to elucidate the possible relations of genetic polymorphisms in C16orf47 gene with serum liver enzyme levels. By examining genotype data of a total of 944 subjects in 5 hospital health promotion center, we discovered the C16orf47 gene polymorphisms are associated with serum liver enzyme levels. The common and highest significant polymorphism was rs7203412 (${\beta}$=3.68, P=3.66E-06) with glutamic oxaloacetic transferase (GOT) and rs7203412 (${\beta}$=6.2, P=7.06E-05) with glutamic pyruvate transaminase (GPT) in all group. Furthermore, the SNP rs7203412 was consistently associated with GOT (${\beta}$=6.41, P=6.78E-08) and GPT (${\beta}$=11.53, P=2.81E-06) in men group. Consequently, we found statistically significant SNP in C16orf47 gene that are associated with serum levels of GOT and GPT. In addition, these results suggest that the individuals with the minor alleles of the SNP in the C16orf47 gene may be more elevated serum liver enzyme levels in the Korean population.

• 대한수의학회지
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• 제44권4호
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• pp.571-579
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• 2004

In order to obtain the genetic informations of the Korean isolates of porcine circovirus 2 (PCV2), nucleotide sequences of total genome of three isolates and open reading frame 2 (ORF2) of four isolates were determined and compared with those of other reference PCV2 isolates. Nucleotide sequences of 3 isolates showed over 99% homology with those of reference strain (GenBank accession no. AF027217). Point mutations were mainly determined on ORF2 regions but little on ORF1 regions. The patterns of pointmutated sites and nucleotide substitution on ORF2 regions were generally consistent between Korean isolates, and these mutated sites observed in Korean isolates were also relatively similar to those of foreign isolates. Phylogenetic analysis of nucleotide or amino acid sequences showed that there were minor branches consisting of three clusters; cluster of Korea, Canada and America, cluster of Spain and Taiwan, and the last cluster of French and China isolates. These results suggested that Korean PCV2s were probably originated from North America such as Canada or USA. The genetic informations obtained from this study could be useful for the research of diagnosis and pathogenecity of PCV2.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.229-233
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• 2010

Pluripotency and self-renewal capacity of human embryonic stem cells (hESCs) are retained by hESCs related genes as OCT4, SOX2 and NANOG. These genes are shown high expression level in diverse cancer cells and have potential role in the carcinogenesis. On the contrary to this, several genes which are up-regulated in the differentiated hESCs are involved to suppress the carcinogenesis or proliferation of cells. We discovered several genes in immortalized lung fibroblast (WI-38 VA13) by suppression subtractive hybridization. Among them, we focused chromosome 6 open reading frame 62 (C6orf62) which is uncharacterized, mapped to 6p22.3 and generated to Hepatitis B virus X-transactivated proteins (HBVx-transactivated proteins, XTP). Aim of this study was to characterize C6orf62 through analyzing of expression pattern in various cell lines. Expression of C6orf62 was significantly upregulated in diverse normal cell lines than cancer cell lines. And C6orf62 was up-regulated in differentiated hESCs (endothelial cells, neural cells) compared to those of undifferentiated hESCs. Also, C6orf62 in WI-38 cells was highly up-regulated during G1/S transition of the cell cycle. Taken together, C6orf62 is shown expression pattern similar to differentiated hESCs-associated genes which down-regulated in cancer cells. Therefore, we assume that C6orf62 may participate to suppress the proliferation and to induce differentiation through regulating the cell cycle.

• Plant Biotechnology Reports
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• 제3권4호
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• pp.309-315
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• 2009

Molecular markers developed from the flanking sequences of two cytoplasmic male sterility (CMS)-associated genes, orf456 and ${\Psi}atp6-2$, have been used for marker-assisted selection of CMS in pepper. However, in practice, the presence of orf456 and ${\Psi}atp6-2$ at substoichiometric levels even in maintainer lines hampers reliable selection of plants containing the CMS gene. In this study, we developed a novel CMS-specific molecular marker, accD-U, for reliable determination of CMS lines in pepper, and used the newly and previously developed markers to determine the cytoplasm types of pepper breeding lines and germplasms. This marker was developed from a deletion in a chloroplast-derived sequence in the mitochondrial genome of a CMS pepper line. CMS pepper lines could be unambiguously determined by presence or absence of the accD-U marker band. Application of orf456, ${\Psi}atp6-2$and accD-U to various pepper breeding lines and germplasms revealed that accD-U is the most reliable CMS selection marker. A wide distribution of orf456, but not ${\Psi}atp6-2$, in germplasms suggests that the pepper cytoplasm containing both orf456 and ${\Psi}atp6-2$ has been selected as CMS cytoplasm from cytoplasm containing only orf456. Furthermore, factors other than orf456 may be required for the regulation of male sterility in pepper.

• International Journal of Industrial Entomology and Biomaterials
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• 제32권2호
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• pp.90-97
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• 2016

The major structural proteins of porcine reproductive and respiratory syndrome virus (PRRSV) are derived from ORFs 4, 5, and 6. They have been considered very important to arouse the humoral and cellular immune responses against PRRSV infection and proposed to be the excellent candidate proteins in the design of PRRS bioengineering vaccine. However, the PRRSV structural proteins are produced in low levels in the infected cells because it forms insoluble protein and possesses several transmembrane regions. To overcome this problem, we fused the ORF4, ORF5, and ORF6 with SUMO (small ubiquitin-related modifier). The resulting fusion protein SUMO-ORF4, -ORF5, and -ORF6 were highly expressed in Bm5 cells. The level of protein expression using the Bombyx mori larvae was higher than that using Bm5 cells. In addition, fusion to SUMOstar, which is not processed by native SUMO proteases, significantly enhanced protein expression levels compared to SUMO fusion. This study demonstrated that SUMO or SUMOstar, when fused with PRRSV structural proteins, was able to promote its soluble expression. This may be a better method to produce PRRSV structural proteins for vaccine development.

• Journal of Microbiology and Biotechnology
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• 제13권3호
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• pp.457-462
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• 2003

The complete nucleotide sequence of a plasmid, pMG1, isolated from Bifidobacterium longum MG1 has been determined. This plasmid, composed of 3,862 base pairs with 65.1% of G+C content. harbors two major open reading frames (ORF) encoding putative proteins of 29 kDa (ORF I) and 71 kDa (ORF II). ORF I showed relatively high amino acid sequence homology with replication proteins of other plasmids from Gr Im-positive and -negative bacteria. Upstream of ORF I, four sets of tandem repeat sequences resembling the iteron structure of related plasmids were found. S1 endonuclease treatment and Southern blot analysis revealed that pMG1 accumulates single-stranded DNA (ssDNA) intermediate, which indicate i the rolling circle replication (RCR) mechanism of this plasmid. Homology search indicated that ORF II encodes plasmid mobilization protein, and the presence of highly conserved oriT sequence in the upstream of this gene supported this assumption. RT-PCR showed that only ORF I is expressed in vivo. Based on these results, pMG 1 was exploited to construct a shuttle vector, pBES2. It was successfully transformed into Bifidobacterium and maintained stably.

• 산업기술연구
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• 제28권B호
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• pp.53-57
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• 2008

From the extensive screening for small cryptic plasmid among about 23 lactic acid bacteria (LAB), 2.4 kb of cryptic plasmid was isolated from Lactobacillus farciminis strain KCTC 3681 and named as pLF24. The plasmid pLF24 was a circular molecule of 2,396 base-pairs in length with a G+C content of 38%. Two protein-coding sequences could be predicted. ORF1 and ORF2 showed homologies to plasmids of gram-positive bacteria. The replication protein coded by ORF2 and the plus origin, were similar to replication regions of other gram-positive bacteria as shown in plasmids such as pLH2, pLS141-1 and pLC2. The nucleotide sequence of pLF24 was deposited into Genbank data base with an accession number of EU429343. The newly isolated plasmid can be used for construction of shuttle vector in Lactobacillus bacteria.

• The Plant Pathology Journal
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• 제34권2호
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• pp.150-156
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• 2018

The genome sequences of two novel monopartite RNA viruses were identified in a common eelgrass (Zostera marina) transcriptome dataset. Sequence comparison and phylogenetic analyses revealed that these two novel viruses belong to the genus Amalgavirus in the family Amalgaviridae. They were named Zostera marina amalgavirus 1 (ZmAV1) and Zostera marina amalgavirus 2 (ZmAV2). Genomes of both ZmAV1 and ZmAV2 contain two overlapping open reading frames (ORFs). ORF1 encodes a putative replication factory matrix-like protein, while ORF2 encodes a RNA-dependent RNA polymerase (RdRp) domain. The fusion protein (ORF1+2) of ORF1 and ORF2, which mediates RNA replication, was produced using the +1 programmed ribosomal frameshifting (PRF) mechanism. The +1 PRF motif sequence, UUU_CGN, which is highly conserved among known amalgaviruses, was also found in ZmAV1 and ZmAV2. Multiple sequence alignment of the ORF1+2 fusion proteins from 24 amalgaviruses revealed that +1 PRF occurred only at three different positions within the 13-amino acid-long segment, which was surrounded by highly conserved regions on both sides. This suggested that the +1 PRF may be constrained by the structure of fusion proteins. Genome sequences of ZmAV1 and ZmAV2, which are the first viruses to be identified in common eelgrass, will serve as useful resources for studying evolution and diversity of amalgaviruses.

• Journal of Microbiology and Biotechnology
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• 제13권5호
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• pp.819-822
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• 2003

Polyketides are an extensive class of secondary metabolites with diverse molecular structures and biological activities. A plasmid-based minimal polyketide synthase (PKS) expression cassette was constructed using a subset of actinorhodin (act) biosynthetic genes (actI-orfl, actI-orf2, actI-orf3, actIII, actⅦ, and actIV) from Streptomyces coelicolor, which specify the construction of an orange-fluorescent anthraquinone product aloesaponarin II, a type II polyketide compound derived from one acetyl coenzyme A and 7 malonyl coenzyme A extender units. This system was designed as an indicator pathway in S. parvulus to generate a hybrid type II polyketide compound via gene-specific replacement. The act ${\beta}-ketoacyl$ synthase unit (actI-orfl and actI-orf2) in the expression cassette was specifically replaced with oxytetracycline ${\beta}-ketoacyl$ synthase otcY-orfl and otcY-orf2). This plasmid-based hybrid PKS cassette generated a novel orange-fluorescent compound structurally different from aloesaponarin II in both S. lividans and S. parvulus. In addition, several additional distinctive blue-fluorescent compounds were detected, when this hybrid PKS cassette was expressed in S. coelicolor B78 (actI-orf2 mutant), implying that the expression of plasmid-based hybrid PKS cassette in Streptomyces species should be an efficient way of generating hybrid type II polyketide compounds.

• 한국응용곤충학회지
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• 제32권3호
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• pp.300-306
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• 1993

Bacillus thuringiensis subsp. kurstaki HD-1으로부터 생산된 살충성 내독소 단백질을 coding하는 CryIIA 유전자를 클로닝하고 염기서열을 조사하였다. HD-1 균주의 12개 plasmid 중 225kb plasmid를 분리하여 CryIIA 유전자를 포함하는 5kb HindIII 절편을 hybridization하여 찾아냈다. 이 절편을 plasmid pUC19에 ligation하여 E. coli에 형질 전환하였다. 이 독소 유전자를 포함하는 4kb BamHI-HindIII 절편은 vector pT7-5에 ligation하여 pSKIIA라하였다. pSKIIA는 3개의 open reading frames(orf1, orf2, orf3)로 구성되어 있으며 염기서열은 3,952base로 되어 있었다. 이러한 3개의 orf 각각의 발현 여부를 확인하기 위하여 생물검정을 하였다. 그러나 orf1 또는 orf2에 의한 형질 전환체는 독성이 없는 것으로 나타났다. orf3를 포함하는 형질 전환체는 3종의 나비목 곤충(배추점나방, 담배거세미나방, 담배나방) 및 1종의 파리목 곤충(집모기) 유충에 대하여 독성을 나타내었다.