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Molecular Characterization of Small Heat Shock Protein(hsp20.8A) from the Silkworm, Bombyx mori
황재삼,구태원,성수일,윤은영,고현정,안미영,김성률,박관호,전재필,강석우,김익수 한국잠사학회 2007 International Journal of Industrial Entomology Vol.15 No.1
To define the molecular mechanism of initiation and termination of diapause during the embryogenesis of silkworms, Bombyx mori, mRNA transcripts from diapausing eggs and diapause activated eggs were compared with differential expression using cDNA array. Among those clones, mRNA transcript from hsp 20.8A, which was expressed at a high level in diapausing eggs that had been incubated at 25oC for 30 days after oviposition, whereas, in the eggs exposed to 15oC for 30 days, 5oC for 60 days, the expression of mRNA decreased. On the other hand, the expression of mRNA during embryogenesis observed abundantly at 4 to 6 day after heat–HCl treatment and later at 9 to 10 day after just before hatching. This result was suggested for us that hsp20.8A was expressed in response to embryogenesis as well as physical stress.
Mariner-Like Elements (MLEs)를 이용한 누에의 분자적 계통 분석
황재삼,이진성,김영섭,성연문 한국생명과학회 1999 생명과학회지 Vol.9 No.2
본 연구는 누에(Bombyx mori)에서 cloning한 BomMAR의 염기서열을 기초로 하여 곤충 MLE(mariner-like ele-ment)의 계통분석을 통한 누에의 분자적 계통 관계를 이해하고자 수행하였다. 전체 10 종의 MLE중에서 15%의 낮은 상동성을 보이는 인간 MLE(Hsmarl)을 제외한 9종의 MLE의 DNA 염기서열을 이용한 UPGMA 분석 결과, BmoMAR을 포함한 10종의 MLE가 세 가지의 subfamily 로 grouping되는 것을 알 수 있었으며, 누에는 genetic distance 0.4332에서 같은 lepidoptera(나비목) 의 H. cecropia, almond moth, webworm 및 microcaddisfly와 함께 grouping 되었다. 따라서 이와 같은 결과는 MLE가 곤충의 계통분석에 유용한 molecular tool이 될 수 있음을 보여준다. In order to understand molecular phylogenetics of silkworm (Bombyx mori), we analyzed the sequences of BmoMAR isolated from Bomhyx mori that is partial coding gene of transposase of mariner-like element(MLE). By pairwise comparing nucleotide sequences of BmoMAR with ten previously reported insect MLEs accessed in GeneBank, the average genetic distance was estimated to be 0.4840. The phylogenetics tree constructed from nine insect species except for human MLE(Hsmarl) by UPGMA method indicated that MLEs are divided into three clusters, and Drosophila mariutiana was independently subgrouped. Bombyx mori(BmoMAR) was subgrouped with microcaddishfly (Orthotrichia cristata), webworm(Atteva punctella), almond moth(Ephestia cautella), Hyalopora cecropia which we lepidoptera. Phylogenetics tree according to UPGMA principle, on the basis of informative nucleotide sequences of nine insect MLEs, indicated that Bombyx mori was more closely related to microcaddishfly(Orthotrichia cristata) and webworm (Atteva punctella) of lepidoptera. We suggest that insect MLEs are a useful key for studying molecular phylogenetics among intra species of insects.
RAPD를 이용한 장려누에품종의 원종간 유전적 유연관계
황재삼,이진성,강현아,이상몽,손해룡 한국응용곤충학회 1997 한국응용곤충학회지 Vol.36 No.3
본 연구에서는 현재 농가에서 일대잡종으로 보급되고 있는 장려누에품종 원종 잠 113 등 20계통에 대해서 RAPD-PRC방법을 이용하여 누에 품봉간 유전적 유연관계를 분석, 검토하였다. 그 결과 공시한 26개의 primer중 24개의 primer에서 다형화 밴드패턴의 마커가 확인되어, 이들 마커를 UPGMA법에 의해 분석하였다. 그 결과, 유전적 유연계수 0.60을 기준으로 하였을 때 2개의 group으로 나눌 수 있었는데 제1group에는 잠113, 잠119, 잠120, 잠123, 잠125와 잠127로서 중국종계잠 120을 제외하고는 모두 일본종계 품종이 포함되어 있었으며, 제 2group은 잠114, 잠121, 잠122, 잠124, 잠126, 잠128, 잠129, 잠130, 잠131, 잠132, 잠133, 잠134, 잠 301과 잠 302로서 일본종계 5품종 중국종계 9품종이 포함되었다. 또한, 잠 114와 잠 120 및 잠114와 잠 127은 유전거리가 다른 품종에 비해서 가장 낮게 분석되었으며, 잠 129와 잠 131은 유전적 유연계수가 1.0으로 두 품종간에는 유전적 유사도가 아주 높았다. The genetic relationships among the twenty parental silkworm, Bombyx mori strains authorized in Korea were evaluated using RAPDs-PCR(Random Amplified Polymorphic DNAs-Polymerase Chain Reaction). Twenty-six different 10-mer oligonucleotide primers were used to screen genetic characteristics of parental twenty silkworm strains by RAPD-PCR analysis. 24 primers showed different banding patterns among the strains. Based on these RAPD patterns, the genetic relationships among the silkworm strains were analyzed by UPGMA(Unweighed Pair-Group Method with Arithmetic average) method. The phylogenetic relationships in the twenty silkworm strains were classified into two major sub-groups at the genetic similarity coefficient of 0.60. The first sub-group included Jaml13, Jaml 19, Jaml20, Jam123, Jam1 25 and Jam 127. Jamll4, Jam1 2 I, Jam 122, Jam 124, Jam1 26, Jam 128, Jam129, Jam 130, Jam 131, Jam1 32, Jam133, Jam134, Jam301 and Jam302 were included in the 2nd group. The genetic distance values among Jam1 14, Jam120 and Jam127 were lower than those among the other strains, while Jam129 is very closely related to Jam131 as showing coefficient value of 1 .O.
RAPD 마커를 이용한 멧누에와 집누에 계통간의 분자적 유연관계 분석
황재삼,이진성,구태원,강현아,손해룡,김호락,Hwang, Jae-Sam,Lee, Jin-Sung,Goo, Tae-Won,Kang, Hyun-Ah,Sohn, Hae-Ryong,Kim, Ho-Rak 한국생명과학회 1998 생명과학회지 Vol.8 No.4
본 연구는 RAPD마커를 이용, 멧누에와 집누에의 분자적 유연관계를 분석하였다. 공시한 35개의 primer에서 166개의 RAPD마커를 얻었으며, 이들 마커를 UPGMA에 의해 분석한 결과, 멧누에와 분자적 유사계수가 가장 낮은 품종은 잠305였고, 가장 높은 품종은 Bibaekjam이었다. 또한, 분자적 유사계수 0.55에서 멧누에와 집누에 계통군으로 분류되고, 0.60에서 3개의 아군 그룹과 2개의 독립개체로 분류되었다. 제1아군에는 J111(일본종계),$pnd^{ps}$(일본종계), Bibaekjam(일본종계)이, 제2아군에는 Galwon(중국종계), C18(중국종계), od yujam JAM306(중국종계), C108(중국종계)이, 제3아군에는 R-hwang(중국종계)이 포함되어 있었고, zebra(유럽종계)와 JAM305(일본종게)는 독립개체로 분류되었다. The molecular relationships have analyzed between the Bombyx mandarina(wild silkworm) and Bombyx mori strains (domesticated silkworm, geographical silkworms). A total of 166 polymorphic RAPD markers amplified from 35 different primers were used to analyze the molecular relationships among thirteen silkworm strains. The genetic similarity coefficient between Bombyx mandarina and Jam305 showed the lowest genetic similarity value with 0.451, Bombyx mandarina and Bibaekjam showed the highest genetic similarity value with 0.958. These strains were classified into Bombyx mandarina(a wild silkworm) and Bombyx mori(twelve domesticated silkworm) groups upon the genetic similary coefficient of 0.55. Further classificient of 0.60; the 1st sub-group (J111, Bibaekjam, $pnd^{ps}$), the 2nd sub-group (Galwon, C18, od yujam, JAM306, C108), the 3rd sub-group(R-hwang, p50), the 4th sub-group(zebra) and the 5th sub-group(JAM305). According to this study, RAPD markers seems to be a valuable tool for molecular relationships and classification among the silkworms.
황재삼,강석우,구태원,고현정,윤은영,최광호,성수일,김성국,이상몽,김익수,김남순 한국잠사학회 2004 International Journal of Industrial Entomology Vol.9 No.1
As an initial step to define the molecular mechanism of diapause during embryogenesis of the silkworm, Bombyx mori, mRNA transcripts from diapausing eggs and diapause-activated eggs were compared by differential expression using cDNA microarray. Twenty-four individual cDNA clones were identified. Amomg them, ten genes including alcohol dehydrogenase, dead box-1, cytochrome oxidase subunit I and 18 wheeler showed increased expression in the diapause-activated eggs. The rest of fourteen genes showed increased expression in diapausing eggs.
황재삼,강현아,이상몽,설광렬,洪性辰,문재유 한국잠사학회 1994 한국잠사곤충학회지 Vol.36 No.2
Differences of feeding response of 280 silkworm genetic stocks and 71 bredding lines to MP-O artificial diet and the mode of the inheritance were invertigated. Feeding response to Mp-O diet varied markedly between the silkworm varieties, and the non-normally distributed curve for the response was observed. From the genetic analysis, the high feeding response to MP-O diet was recessive to the low feeding response. Therefore it is considered that the high feeding response of the newly hatched silkworm larvae to MP-O diet is controlled by a recessive gene.
황재삼,구태원,성수일,윤은영,강석우,김익수,최광호,김근영,이상몽 한국잠사학회 2004 International Journal of Industrial Entomology Vol.9 No.2
We constructed an oligo-d(T) primed directional cDNA library from the Bombyx mandarina whole larvae. In an effort to isolate genes expressed in the B.mandarina, 227 expressed sequence tags (ESTs) were generated by single-pass sequencing from the cDNA library. Sequence analysis showed that 107 clones (47.1%) were classified into known genes and 120 clones (52.9%) were novel transcripts, which are unknown for their function. Of the 107 known genes, the most abundant gene was found to be actin and followed by serine protease in the expression profile. Among these clones, a serine protease homolog (BmSP) which is a class of proteolytic enzymes isolated. Full-length sequence of the BmSP cDNA clone was 922 bp in length and has an open eading frame of 276 amino acids. The conserved histidine, aspatic acid and serine resihues forming the catalytic center as well as cysteine residues contributing to three disulphide bonds also were found in Bmsp ene. mRNA expression analysis revealed a high and specific expression of the gene only in midgut tissue, suggesting that BmSP gene is closely associated with the expression of digestive enzyme.