Optimization of Parameters for GUS Gene Transformation of Porphyra yezoensis by Particle

남보혜,홍용기,박중연,진덕희 한국수산과학회 2006 Fisheries and Aquatic Sciences Vol.9 No.4

지질다당질로 자극한 넙치 백혈구의 발현유전자의 분석

남보혜,문지영,김영옥,김우진,공희정,이상준,최태진,Nam, Bo-Hye,Moon, Ji-Young,Kim, Young-Ok,Kim, Woo Jin,Kong, Hee Jeong,Lee, Sang-Jun,Choi, Tae-Jin 한국해양바이오학회 2007 한국해양바이오학회지 Vol.2 No.3

넙치의 생체방어 및 면역관련 유전자의 대량확보를 위하여, 지질다당질로 자극한 넙치의 말초백혈구 cDNA library를 suppression subtractive hybridization 방법으로 제작하여 발현유전자의 발현 양상 및 특성을 분석하였다. 총 470개의 SSH 클론 중 기존에 밝혀진 유전자와 상동성을 보인 218개의 클론을 분석한 결과, 대표적인 전염증사이토카인 IL-1의 유인수용체인 IL-1RII가 34번의 가장 높은 빈도로 발현되었으며, IL-$1{\beta}$가 14번의 높은 발현 빈도를 보였다. 그 중 펩티도글리칸 인식 단백질및 혈관활성 장 펩타이드 유전자는 어류에서는 처음으로 보고되는 것으로, 지질다당질로 자극한 넙치의 말초백혈구 SSH cDNA library는 다양한 생체방어 및 면역관련 유전자를 포함하고 있었다. Suppression subtractive hybridization (SSH) was performed to construct an cDNA library of olive flounder Paralichthys olivaceus peripheral blood leukocytes (PBL) stimulated with lipopolysaccharide (LPS). Total 470 clones were randomly selected and sequenced, 214 out of 470 sequences showed identities with known genes and 252 sequences were unknown genes. Among the 218 known genes, 34 sequences were found to be homologous to IL-1RII gene, and 14 sequences were identified as IL-$1{\beta}$. RT-PCR analysis showed that IL-$1{\beta}$ mRNA was induced 1h after LPS-stimulation, and IL-1RII was increased from 3h after stimulation, indicating that most of SSH clones are associated with inflammatory responses in fish, and this SSH cDNA library would be useful to identify bio-defense and immuno-related genes in fish.

유전적 다양성 분석을 활용한 꼬막의 원산지 판별

남보혜,동춘매,이미난,김은미,김영옥,노은수 한국패류학회 2024 The Korean Journal of Malacology Vol.40 No.1

This study aimed to develop a genetic analysis method to identify the origin of cockles (Tegilarca granosa). Initially, analysis of the mitochondrial COI gene revealed genetic variations among Korean, Chinese, and Japanese cockles. While distinguishing between Chinese and Japanese cockles was challenging, a specific haplotype (H1) found only in Korean cockles allowed researchers to infer their origin. Next, we analyzed the genetic characteristics of cockle populations using 19 microsatellite markers. This analysis validated the accuracy of assigning a cockle's origin and assessed the reliability of this method. The analysis revealed that three genetic loci (Teg02, Teg08, TMP18) displayed differing numbers of alternative alleles expressed between domestic populations and Chinese and Japanese populations, indicating genetic differentiation due to geographic separation. Using 16 microsatellite markers, the accuracy and reliability of origin determination achieved 89.6% and 86.6%, respectively. This shows there's enough information to distinguish between Korean, Chinese, and Japanese cockles. The results of this study demonstrate the usefulness of genetic analysis methods in determining the origin of cockles. This method is expected to contribute significantly to ensuring the safety and quality management of cockle distribution in the future. Additionally, there is a need for continued research to improve the accuracy and ability to differentiate origins through further development of microsatellite marker-based methods.

Analysis of heat, cold or salinity stress-inducible genes in the Pacific abalone, Haliotis discus hannai by suppression subtractive hybridization

남보혜,박은미,김영옥,김동균,이영주,이상준,안철민 한국패류학회 2013 The Korean Journal of Malacology Vol.29 No.3

In order to investigate environmental stress inducible genes in abalone, we analyzed differentially expressed transcripts from a Pacific abalone, Haliotis discus hannai, after exposure to heat-, cold- or hyposalinity-shock by suppression subtractive hybridization (SSH) method. 1,074 unique sequences from SSH libraries were composed to 115 clusters and 986 singletons, the overall redundancy of the library was 16.3%. From the BLAST search, of the 1,316 ESTs, 998 ESTs (75.8%) were identified as known genes, but 318 clones (24.2%) did not match to any previously described genes. From the comparison results of ESTs pattern of three SSH cDNA libraries, the most abundant EST was different in each SSH library: small heat shock protein p26 (sHSP26) in heat-shock, trypsinogen 2 in cold-shock, and actin in hyposalinity SSH cDNA library. Based on sequence similarities, several response-to-stress genes such as heat shock proteins (HSPs) were identified commonly from the abalone SSH libraries. HSP70 gene was induced by environmental stress regardless of temperature-shock or salinity-stress, while the increase of sHSP26 mRNA expression was not detected in cold-shock but in heat-shock condition. These results suggest that the suppression subtractive hybridization method is an efficient way to isolate differentially expressed gene from the invertebrate environmental stress-response transcriptome.

Characterization of the Heat Shock Protein 60 and 10 Genes andtheir Bidirectional Promoter Region in Olive Flounder (Paralichthysolivaceus)

남보혜,임영옥,임상욱,조용철,김봉석,지영주,이상준,정재훈 한국유전학회 2006 Genes & Genomics Vol.28 No.4

Heat shock protein (HSP) 60 and 10 are stress-inducible mitochondrial matrix proteins that form an important chaperonin complex for mitochondrial protein folding and function. Extensive studies on model fish species have revealed molecular chaperones, such as HSP90, HSP70, and low molecular weight HSP (16–47 kDa), but the study of chaperonins in fish has not yet been conducted. We present the full sequence of the HSP60 and HSP10 genes for the teleost olive flounder (Paralichthys olivaceus), which were found to be linked head-to-head comprising approximately 10 kb and consisting of 11 and 4 exons, respectively. Using a luciferase–reporter assay, we demonstrated that the region between the two genes functions as a bidirectional promoter. The nucleotide variations detected are localized in a putative CCAAT-box in the bidirectional promoter region, and analysis of the transcriptional activity of the promoter fragment with these variations showed that it affects transcriptional levels.

Quantitative viability of seaweed tissues assessed with 2,3,5-triphenyltetrazoli

남보혜 부경대학교 수산과학연구소 1998 硏究業績集 Vol.23 No.-

Quantitative viability of seaweed tissues assessed with 2,3,5-triphenyltetrazoli

남보혜 부경대학교 기초과학연구소 1998 부경대 기초과학연구논문집 Vol.1 No.-

EST-based Survey of Gene Expression in Seven Tissue Types from the Abalone Haliotis discus hannai

박은미,남보혜,김영옥,공희정,김우진,이상준,공인수,최태진 한국수산과학회 2007 Fisheries and Aquatic Sciences Vol.10 No.3

Genetic Variation in Wild and Cultured Populations of the Sea Squirt Halocynthia roretzi Inferred from Microsatellite DNA Analysis

한현섭,남보혜,강정하,김이경,지영주,허영백,윤문근 한국수산과학회 2012 Fisheries and Aquatic Sciences Vol.15 No.2

We used nine microsatellite DNA markers to estimate genetic variation among wild and cultured populations of the sea squirt Halocynthia roretzi. The loci were polymorphic, with 6-32 alleles, and allelic richness ranged from 6.0 to 26.1 in each population. The wild and the cultured populations had similar mean heterozygosities (HO and HE), allele numbers, and allelic richness. One cultured population with softness syndrome had a lower mean in the observed heterozygosity (HO = 0.57) and higher mean inbreeding coefficient (FIS = 0.261) than any other populations. This suggests that the loss of genetic variation in the diseased population might be due to increased inbreeding. A neighbor-joining tree and pairwise population estimates of FST showed moderate genetic differentiation between the wild and the cultured populations. Additionally, the softness syndrome population was genetically divergent from wild populations, but it was genetically close to the cultured populations.

북방전복, Haliotis discus hannai 3배체의 생식능력

지영주,남보혜,이정용,장영진 한국패류학회 2013 The Korean Journal of Malacology Vol.29 No.2

참전복의 배수체별 생식소 조직 관찰에서 2배체 난소의 경우에는 성숙기를 거쳐 방출기로 들어가고 있는 상태이며, 2배체 정소의 경우에는 방출기의 상태였다, 이에 비해 3배체 전복의 생식소는 발달이 억제되어 난소는 성장기 혹은 성숙기의 상태이고, 정소는 성숙기의 단계에 머물고 있다. 따라서 전복은완전한 불임이 되는 것은 아니고 어느 정도 성숙이 억제된다고생각된다. 3배체 참전복의 산란 유발 실험에서 산란 유발에 반응한 개체는 2배체는 암수 전부 반응해서 100%의 산란 유발율을 나타냈고, 3배체는 암컷 50%, 수컷은 25%의 산란 유발율을 나타냈다. 산란된 알과 정자를 배수체별로 조사한 결과, 2배체의 정자는 장경 17.47 μm, 단경 11.40 μm 및 정자의 길이103.94 μm이었던 데 비해 3배체 정자는 장경 10.86μm, 단경 8.47 μm 및 정자의 길이 85.06 μm로서 2배체 정자가 3배체 정자보다 장경, 단경, 정자의 길이 모두에서 유의적으로길었다 (P < 0.05). 알의 직경은 2배체가 199.50 μm, 3배체가 192.50 μm로서 2배체가 컸으나 유의한 차는 없었다( P > 0.05). 배수체별 교배 실험에서 2배체 × 2배체는 정상적으로 수정되어서 발생을 했으나 2배체 알 × 3배체 정자와 3배체 알 ×2배체 정자의 경우에는 수정이 되지 않았다. The gonadal development of triploid and diploid Pacific abalones, Haliotis discus hannai was histologically investigated in spawning season. Diploid abalones had matured oocytes and spermatozoa, but most triploid had spermatocytes or developing oocytes that was slightly retarded in gonadal development compared to diploid abalones. In spawning experiment of triploid and diploid abalones, spawning rates of diploid male and female were 100%, but those of triploid female was 50% and male was 25% respectively. Investigation of spawned abalone eggs and spermatozoa revealed that length of diploid sperms head were 17.47 μm, breadth of head were 10.31 μm, length of spermatozoa were 130.72 μm, but those of triploid spermatozoa were 11.83 μm, 7.89 μm and 103.36 μm respectively. Triploid spermatozoa were significantly small to diploid spermatozoa (p < 0.05). The eggs of diploid and triploid were not different in size. The cross experiment between oocytes produced by triploids and spermatozoa by diploids (3n × 2n cross) revealed that no fertilization were occurred, and 2n × 3n cross also revealed same result.