멧누에 ( Bombyx mandarina ) 난간단백질 유전자의 유전적 분석

성승현(Seung Hyun Sung),김순정(Soon Jeung Kim),염승식(Seung Shic Yum),김종길(Jong Kil Kim),김기세(Ki Sei Kim),김근종(Keun Chong Kim),문준옥(Jun Ok Moon),이병순(Byong Soon Lee),서동상(Dong Sang Suh) 한국유전학회 2001 Genes & Genomics Vol.23 No.4

Two multigene families, high-cysteine A(HcA) and high-cysteine B(HcB), are located within 140 kb region of chorion locus in Bombyx mori. Each family consists of 15 members and one of each members are transcribed in pairs. We report isolation and characterization of HcA and HcB genes of the wild type silkmoth, Bombyx mandarina. In this study, we examined 6 repeating units out of 15 HcA genes and 6 units of 15 HcB genes of B. mori and B. mandarina, respectively, and found that the arrangements and expression units of these are dramatically different. In addition, we found that most of the sequence variations were located within the C-variable region. These results suggest that the likely mechanism of concerted evolution is gene conversion-like event in HcA/HcB gene pairs.

멧누에 난각단백질 유전자(Hc)의 클로닝 및 구조해석

김종길,성승현(Seung Hyun Sung),남혜영,최지영,김삼은,(T. Amura),박영민,서동상(Dong Sang Suh) 한국잠사학회 2000 한국잠사학회 잠사과학 100주년 국제심포지엄 및 추계학술대회 Vol.- No.-

마우스 Sp2/0 세포의 형질전환에 관한 연구

손정필,성승현,서동상 성균관대학교 생명과학자원연구소 1995 生命資源科學硏究 Vol.2 No.1

Transfection, gene transfer into mammalian cells, is used to study gene structure and function. In this study, transfection of Sp2/0 cells using LK4LAC plasmid DNA(containing β-galactosidase gene) with various parameters was investigated. The transformants were identified by the β-galactosidase staning method. Optimal transfection conditions of Sp2/0-Ag14 cells were studied with two methods, calcium phosphate(CaPO_4) precipitation method and liposome method. In calcium phosphate precipitation method, the highest transfection efficiency was obtained when 20㎍ of DNA was transfected for 12hours and incubated for 16hours. In case of liposome method, the optimal. condition was achieved when 25㎍ of lipofectin amount was transfected for 18hrs. The optimal drug concentration of selection with G418 was 400㎍/㎖ in the 20 cells/dish of Sp2/0-Ag14 cell line.

누에 미토콘드리아 유전체의 제한효소 지도작성, 클로닝 및 염기서열 분석

이진성,성승현,김용성,서동상 한국잠사학회 2000 한국잠사곤충학회지 Vol.42 No.1

The mitochondrial genome of domesticated silkworm (Bombyz mort) was mapped with five restriction endonucleases (BamHI, EcoRI, HrndIII, PstI and Xbal), the entire genome was cloned with HindIII and EcoRI. From the end sequencing results of 5` and 3` region for full genoe set of eleven mitochondrial clones, the seven mitochondrial genes(NADH dehydrogenase 6, ATPase 6, ATPase 8, tRNALys, tRNAASP. tRNAThr and tRNAPhe ) of mori were identified on the basis of their nucleotide sequence homology. The nucleotide composition of NADH dehydrogenase 6 was heavily biased towards adenine and thymine, which accounted for 87.76%. On basis of the sequence similarity with published tRNA genes from six insect species, the tRNALys, tRNAASP. tRNAThr were showed stable canonical clover-leaf tRNA structures with acceptable anticodons. However, both the DHU and TΨC arms of tRNAPhe could not form any stable stem-loop structure. The two overlapping gene paris (tRNALys-tRNAASP and ATPase8-ATPase6) were foud from our sequencing results. The genes are encoded on the same strand. ATPase8 and ATPase6 overlaps(ATGATAA) which are a single example of overlapping events between abutted protein-coding genes are common. and there is evidence that ATPase8 and at an internal start site for ATPase6. Ultimately, this result will provide assistance in designing oligo-nucleotides for PCR amplification, and sequencing the specific mitochondrial genes for phylogenetics of geographic races, genetically improved silkworm strains and wild silkworm (mankarina) which is estimated as ancestral of domesticated silkworm.

미세주사법에 의한 P 인자를 벡터로 이용한 송사리의 형질전환에 관한 연구

김남철,성승현,서동상 성균관대학교 생명과학자원연구소 1996 生命資源科學硏究 Vol.2 No.2

어류의 형질전환 실험에 있어서의 두 가지 문제점은 어류의 수정란마다 독특한 특성을 갖고 있어 독특한 미세주입법이 요구된다는 점과 적절한 벡터계가 구축되어 있지 않다는 것이다. 본 연구는 어류의 형질전환에서 문제가 되는 난각의 단단함을 극복하는 미세주입법의 확립과 P벡터계의 유용성의 검증에 필요한 미세주입된 벡터의 생체 내 존재 여부를 검출하기 위한 방법조건 확립이라는 두 가지 목적을 가지고 수행하였다. 먼저 미세주입법에 있어서는 5% agar 평판을 지지체로 사용하여 바늘과 수정란을 동시에 고정시킴으로써 미세주입법을 수행할 수 있었으며 17.3%의 생존율을 보였다. 두 번째로 미세 주입이 되고 부화된 개체에 실제로 P벡터의 효과적인 미세주입이 수행되었고 생체 내에 어떤 방식으로든 그 벡터가 존재하는가를 검출하기 위해서는 PCR을 이용하였다. P벡터의 역반복 서열을 primer로 사용한 결과 5.1Kb의 증폭 산물이 관찰되었다. 이 두 방법은 앞으로 P벡터께의 유용성을 점진적으로 검증해 나가는데 유용하게 이용될 수 있을 것이다.

RAPD-PCR을 이용한 마우스 계통간 유연관계

강은영,성승현,이진성,김선호,서동상 성균관대학교 생명과학자원연구소 1996 生命資源科學硏究 Vol.3 No.2

We have analyzed the genetic relationship among the mouse strains using RAPD-PCR. Polymorphic RAPD markers amplified from various primers were used to analyze the genetic relationship among seven mouse strains, CBA, C57BL/6, BALB/C, NOD, A/wy, DBA2, and AKR. The genetic similarity coefficients were estimated using the RAPD markers by UPGMA method. The genetic similarity coefficient between CBA and NOD was 0.474. This value was lower than any other genetic similarity coefficient among the strains tested. And CBA-specific RAPD band patterns were observed in 5 primers from 6 polymorphic primers used. This study makes possible phylogenetic identification among the inbred mouse strains at the molecular level using RAPD-PCR techniques.

멧누에 난각유전자의 구조해석 및 진화관계 연구

김기세,성승현,김순정,서동상 성균관대학교 생명공학연구소 1999 生命工學硏究 Vol.5 No.1

The chorion protein (A, B, C, D, E, HcA, HcB) are synthesized by monolayer of follicular epitherial cells and assembled after secretion into eggshell in B. mori. The chorion gene contains 15-multigene families (the pair-HcA, HcB) which are divergently oriented gene pairs in B. mori. The chorion protein of B. mandarina (Korean wild silkmoth) apparently differ those of B. mori in the structure, morphology and protein secondary structure. The chorion gene of B. mandarina have few been studied yet. We've studied the chorion gene of B. mandarina, constructed wild silkmoth genomic library and obtained the 10 Hc clones. 8 clones are sequenced and characterized (some are partial sequence). The sequence similarities of B. mori and B. mandarina of each families are 90% and 85% respectively. Some of them are changed the charge of amino acids in the terminal of central domain. The variable region of C-arm is composed of Cys-Gly, Cys-Gly-Gly repeated structure which determines the length variation in B. mandarina.. It means the genetic exchanges are occurred in the region among multigene families. We could find the structural and evolutionary relationship.

노랑초파리 한국 자연집단 내의 체중에 관한 유전 분산 분석

김용성,박영민,서동상,성승현,심미정,성기창,김근종 한국유전학회 1998 Genes & Genomics Vol.20 No.3

We estimated genetic variance components for body weight in D. melanogaster. The second chromosomes of 98 lines were extracted from a Korean natural population. Analyses of heritability and genetic variance were performed by partial diallelic crosses. The correlation coefficient (r) of body weight between females and males was estimated to be 0.2817, which was highly significant at the 0.1% significance level. The heritability (H²) was estimated to be 0.317 for females and 0.308 for males ; the additive genetic variance (σ²A) 0.00136±0.00096 and 0.00127±0.00047; the dominance variance (σ²D) -0.00014±0.00031 and -0.00052±0.00026, respectively. These data indicate that, in this population, mutation-selection balance might be an explanation for the mechanism for the maintenance of genetic variability of body weight.

누에의 RFLP marker 개발

김용성,이진성,황재삼,이상몽,성승현,서동상 한국유전학회 1999 Genes & Genomics Vol.21 No.4

To construct RFLP linkage map of silkworm (Bombyx mori), as a preliminary step, we assessed the efficiency for single or low copy sequences from silkworm genomic DNA. A genomic DNA plasmid library was constructed using EcoRl-digested 0.3 to 5 kb DNA fragments of II-111 strain. As results of two step screening (reverse colony hybridization and reverse southern hybridization) using a total genomic DNA as a probe, about 6% (70/1,200) of clones examined was estimated single or low copy sequences. At least 80% of total clones contained middle or high repetitive sequences. These results indicate that B. mori genome has a lot of high repetitive sequences. However, analysis for its precise copy number in the genome remains to be elucidated. The obtained 70 single-sequences will be useful molecular markers to study silkworm genomic analysis in the near future, although it has low efficiency of single copy sequence for constructing linkage map.

열처리 반응에 대한 노랑초파리의 유전적 변이

김용성,서동상,박정훈,성승현 한국유전학회 1997 Genes & Genomics Vol.19 No.3

Adult flies were collected from Suwon population and the third chromosomes were extracted from each of 112 males using the Ser/Sb balancer stocks. Then the homozygote viabilities of the third chromosome were estimated under the normal (25℃) and the heatshock (37℃) conditions. A degree of response(name as Responsibility) was measured by the ratio of viabilities under the heat shock and the normal condition. Furthermore, the puffing patterns of the strains which have shown high and low responsibility were analyzed with heat-induced puffs on the polytene chromosome in order to observe relationship between responsibility and the expression of Hsp genes. The following results were obtained. 1) The lethal frequency on the third chromosome of homozygous strains was 0.277+0.042.2) The average viabilities of non-lethal homozygous strains were estimated 0.800+0.026 and 0.726+0.026 at normal and heat shock condition, respectively. 3) The average responsibility was estimated 0.938+0.032. The wide distribution of responsibilities represented that there were many of variants for heat-shock responses in the third chromosome of the natural population. 4) Comparing with the puffing patterns on the polytene chromosome, it was observed that the duration of the puffs of 87A and 87C, which were known to code Hsp70, was longer in the strains of high responsibility than in those of low one.