• Animal cells and systems
• /
• 제9권3호
• /
• pp.107-111
• /
• 2005

C. elegans DLK-1 has been reported to play an important role in synaptogenesis by shaping the structure of presynaptic terminal. In this study, we investigated the expression pattern and regulation of the dlk-1 gene in C. elegans. To determine the expression pattern, we made a dlk-1::gfp fusion construct, named pPDdg1, which consisted of -2.2 kb 5' upstream region, the first exon, the first intron, and a part of the second exon of the dlk-1 gene. By microinjecting this construct into the worm, we observed that the DLK-1::GFP was expressed mainly in neurons. We next examined the regulatory elements of gene expression by deletion analysis of pPDdg1. Removal of a large portion of the 5' upstream region (${\Delta}-361$ to -2246) of the gene had little effect on the expression pattern, whereas deletion of the first intron led to elimination of the DLK-1::GFP expression in most of the neurons. Our results suggest that the first intron of the C. elegans dlk-1 gene contains the regulatory element critical for gene expression.

• Fisheries and Aquatic Sciences
• /
• 제14권4호
• /
• pp.303-310
• /
• 2011

We characterized a cytoplasmic actin gene locus in greenling Hexagrammos otakii (Scorpaeniformes). Genomic clones isolated from the greenling DNA library contained two homologous cytoplasmic actin gene copies (HObact2.1 and HObact2.2) in a tail-to-head orientation. Their gene structure is characterized by six translated exons and one non-translated exon. Exon-intron organization and the nucleotide sequences of the two actin gene isoforms are very similar. However, only the HObact2.1 isoform contains microsatellite-like, dinucleotide repeats in the 5'-flanking region (named HOms2002) and intron 1 following the non-translated exon 1 (named HOms769). One microsatellite locus (HOms769) was highly polymorphic while the other (HOms2002) was not. Based on bioinformatic analysis, different transcription factor binding motifs are related to stress and immune responses in the two actin isoforms. Semiquantitative and real-time reverse transcription-PCR assays showed that both isoform transcripts were detectable ubiquitously in all the tissues examined. However, the basal expression levels of each isoform varied across tissues. Overall, the two isoforms showed a similar, but not identical, expression pattern. Our data suggest that the cytoplasmic actin genes may be the result of a recent duplication event in the greenling genome, which has not experienced significant subfunctionalization in their housekeeping roles.

• 생명과학회지
• /
• 제14권6호
• /
• pp.1009-1017
• /
• 2004

인간유래 시알산전이효소 유전자들의 특이적 발현과 그들의 mRNA isoform의 생성에 대한 조절기구를 이해하기 위하여 5종류의 human 시알산전이효소 유전자(hST3Cal II, hST8Sia II, hST8Sia III, hSTS8Sia IV, hST8Sia V)들의 게놈구조를 분석하였다. hST3Gal II 유전자는 17 kb이상의 게놈상에 46 bp에서 1017 bp의 길이를 가진 exon이 6개로 이루어져 있고, hST8Sia III유전자는 10 kb이상의 게놈상에 125bp에서 2023bp의 길이를 가진 exon이 4개로 이루어져 있어 다른 human 시알산전이효소 유전자들보다 짧고 단순한 구조를 가지고 있었다. 반면에 다른 3종류의 유전자(hST8Sia II, hST8Sia IV, hST8Sia V)들은 70 kb이상의 게놈상에 5개이상의 exon으로 이루어져 있으며, 5종류 모두 exon-intron boundary는 GT-AG rule을 나타내고 있었다. 특히 모든 시알산전이효소에 고도로 보존되어 있는 sialylmotif L은 hST8Sia III유전자에서는 하나의 exon에 존재하는 반면에, 다른 시알산전이효소 유전자에서는 분리된 exon에 존재하여 exon의 구조적 다양성을 나타내고 있다. 또한, 본 연구에서는 5'-RACE와 cap site hunting법에 의해 hST3Gal II 유전자의 전사개시점을 결정하였다.

• Journal of Animal Science and Technology
• /
• 제45권2호
• /
• pp.169-182
• /
• 2003

ADRP 유전자가 24개월령 한우 등심조직에서 발현량이 급격히 증가하여 30개월령 등심조직에서는 발현량이 다소 감소하는 발현양상 분석결과로부터 이전 연구에서는 ADRP 유전자를 한우 성장단계 특이발현 유전자로 선정하였다. 본 연구에서는 ADRP 유전자의 발현조절 기작을 분석하기 위하여 promoter 영역을 포함하는 ADRP 유전자 전체영역을 cloning하였으며, 구조를 분석하고 promoter의 특성을 조사하였다. 한우 ADRP cDNA 단편을 probe로 합성하여 Southern blot 분석을 실시한 결과로부터 ADRP 유전자가 한우 genome 상에서 single copy로 존재하고 크기는 대략 12 kb에 해당하는 것을 확인하였다. Genomic DNA library screening을 실시하여 promoter 영역을 포함하는 ADRP 전체 유전자에 해당하는 clone을 확보하고 HwADRPg-1으로 명명한 후, 염기서열을 결정하고 분석하였다. 한우 ADRP 유전자, HwADRPg-1은 8개의 exon과 7개의 intron으로 구성되어 있으며 모든 exon-intron 경계는 GT/AG 원칙을 따르고 있었고, coding 영역은 7,633 bp로서 6개의 intron에 의해 7개의 exon으로 나누어져 있었다. HwADRPg-1의 promoter 영역에서는 TATAA box는 발견되지 않았으며, -70 위치에 근육 특이적 transcription activator인 Myo G 서열이 존재하였고, -629 위치에는 지방세포의 분화를 유도하는 것으로 알려진 C/EBP (CCAAT/enhancer binding protein) 서열이 존재하였다. HwADRPg-1의 조절영역에 있는 Myo G factor가 근육조직에서 ADRP 유전자가 발현될 수 있도록 하며, 근육의 발달정도를 신호로써 감지하여 근육조직에서 성장단계에 따른 ADRP 유전자의 발현량을 조절할 것으로 추정되고, 다른 종류의 지방세포 특이적인 전사인자 및 지방세포의 분화정도를 신호로 인식하는 전사단계 조절인자를 조사하기 위하여 promoter 영역의 추가분석이 이루어져야 할 것으로 사료된다.

• Journal of Plant Biology
• /
• 제38권2호
• /
• 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.

• Asian-Australasian Journal of Animal Sciences
• /
• 제12권5호
• /
• pp.689-694
• /
• 1999

This experiment was carried out to isolate the partial bovine (Bos Taurus coreanae) myogenic factor encoding gene, MyoD, using the rat myogenic factor (MyoD) gene sequence and to compare the gene sequence between another myogenic factor (Myf 5) and MyoD gene of the bovine. To make the probe and isolate the MyoD gene, PCR was performed to amplify rat and bovine MyoD gene including exon I, II and intron I. The homology between mouse and bovine MyoD is high; bovine MyoD gene shows 17 different gene sequence region compared to rat MyoD. Among those, two regions have significant differences; one is the exon I part between 2834 and 2850 bp, the other is intron part between 3274 and 3303 bp of the mouse. At this region homology was 40% in the former and 50% in the latter. Homology between bovine MyoD and Myf5 was 83% in the exon 1. Especially exon I in the Myf5 602-617 bp and 651-683 bp have significant differences. These results suggest that MyoD gene have a similar gene structure in mouse and bovine and MyoD and Myf5 of the bovine, at least in part, have a similar expression and activity.

• 대한화학회지
• /
• 제51권6호
• /
• pp.543-548
• /
• 2007

MODY는 일반적으로 제 2 형 당뇨의 특수형으로 25세 이전에 발병하며 상염색체 우성으로 유전되 며 인슐린 분비장애를 특징으로 한다. GCK와 HNF-1α의 유전자 돌연변이는 제 2 형 당뇨에서 가장 큰 원인 중에 하나이다. 따라서 이들 유전자다형성의 연관성을 다양한 분석 방법으로 연구할 필요성이 대두되고 있다. GCK와 HNF-1α 유전자의 exon과 exon에 근접한 intron을 실험하였고, 또한 promotor까지 PCR-DHPLC (Polymerase Chain Reaction - Denaturing High Performance Liquid Chromatography) 방법과 direct sequencing 방 법을 사용하였다. 시료는 11명의 환자와 20명의 정상인에서 DNA를 추출하였고 GCK와 HNF-1α의 단일 염기 다형성을 PCR-DHPLC 방법으로 확인하였다. 결과적으로 GCK 유전자에서는 1개(R135G)를 검출하였고 HNF-1α 유전자에서는 2개(I27L, S487N)를 검출하였으며 intron 8에서도 돌연변이를 검출할 수 있었다.

• Journal of Animal Science and Technology
• /
• 제49권3호
• /
• pp.295-302
• /
• 2007

본 연구는 한우 24두의 혈액으로부터 geno- mic DNA를 추출하여 leptin 유전자의 intron 1부터 3’-UTR 부분까지의 염기 서열을 결정하여 총 25개의 SNPs을 발굴하였다. 발굴된 SNPs 중 16개는 기존의 서양 품종에서도 발견되는 것으로 그 빈도는 대부분의 경우 유사하였으나, 서로 유의성이 있도록 상이한 경우도 있었다. Intron 1의 T1064G, exon 2에서 발견된 nonsynonymous SNP(T1163A, Val to Glu)과 exon 3의 C3011G 및 G3256A(Gly to Asp) SNPs은 기존에 보고되지 않은 한우에서 새롭게 발견된 것으로 종 간의 차이를 나타내는 것으로 사료된다. 이러한 한우의 SNPs 정보는 한우의 유전형을 결정하고, 도체 및 육질 형질 등 중요 경제 형질과의 연관 분석을 통하여 중요한 유전자 표지 확보 및 한우 판별 등의 응용에 이용될 수 있을 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제24권7호
• /
• pp.898-904
• /
• 2011

The PCR- restriction fragment length polymorphism (RFLP) in and around TM4 of SLC11A1 gene and its association with the incidences of brucellosis in Hariana breed (Bos indicus) and Holstein Friesian crossbred (Bos indicus${\times}$Bos taurus) cattle was examined. A fragment of 954 bp encoding the TM4 was amplified, and RFLP was identified by digestion of the amplicon independently with AluI and TaqI. The amplicon (GenBank Acc. No. AY338470 and AY338471) comprised of a part of exon V (<59 bp) and VII (62>), and entire intron 5 (423 bp), exon VI (71 bp) and intron 6 (339 bp). Digestion with AluI revealed the presence of two alleles viz, A (281, 255, 79 and 51 bp) and B (541, 255, 79 and 51 bp). The frequency of A allele was estimated as 0.80 and 0.73 in Hariana and crossbred cattle, respectively. Due to presence of a polymorphic TaqI site at intron 5, two alleles: T (552 and 402 bp) and Q (231, 321 and 402 bp) were identified. The frequency of T allele was estimated as 0.96 and 0.97, respectively. For association study, on the basis of serological tests and history of abortion, the animals were grouped into "affected" and "non-affected". However, no association could be established with the observed RFLPs.

• Asian-Australasian Journal of Animal Sciences
• /
• 제27권9호
• /
• pp.1228-1235
• /
• 2014

The swine leukocyte antigen (SLA)-DRA locus is noteworthy among other SLA class II loci for its limited variation and has not been investigated in depth. This study was investigated to detect polymorphisms of four exons of SLA-DRA gene and its association with piglet diarrhea in Landrace, Large White and Duroc pigs. No polymorphisms were detected in exon 3, while 2 SNPs (c.178G>A and c.211T>C), 2 SNPs (c.3093A>C and c.3104C>T) and 5 SNPs (c.4167A>G, c.4184A>G, c.4194A>G, c.4246A>G and c.4293G>A) were detected in exon 1, exon 2 and exon 4 respectively, and 1 SNP (c.4081T>C) in intron 3. Statistical results showed that genotype had significant effect on piglet diarrhea, individuals with genotype BC had a higher diarrhea score when compared with the genotypes AA, AB, AC and CC. Futhermore, genotype AC had a higher diarrhea score than the genotype CC in exon 1 (p<0.05); diarrhea scores of genotype AA and BB were higher than those of genotypes AC and CC in exon 2 (p<0.05); individuals with genotype AA had a higher diarrhea score than individuals with genotype AB and BB in exon 4 (p<0.05). Fourteen common haplotypes were founded by haplotype constructing of all SNPs in the three exons, its association with piglet diarrhea appeared that Hap2, 5, 8, 10, and 14 may be the susceptible haplotypes and Hap9 may be the resistant haplotype to piglet diarrhea. The genetic variations identified of the SLA-DRA gene may potentially be functional mutations related to piglet diarrhea.