콩(Glycine max)의 공생균주 Bradyrhizobium japonicum SNU001의 특성

고세리,박용근,안정선 한국미생물학회 1991 미생물학회지 Vol.29 No.2

The root nodules and Glycine max were classified as determinate nodule based on their morphological characteristics, and isolated endosymbiont as a Bradyrhizobium based on its growth rate and single subpolar flagellum. The isolate was similar to B. japonicum USDA110 in utilization of carbon source, growth at 38.deg.C and 2% NaCl, production of $H_{2}$S and especially in the restriction endonuclease digestion pattern of symbiotic genes, allowing them to be placed in sTI group together. The former, however, grew better than the later in broad pH range from 5.0 to 9.5. Infectivity and effectivity of the isolate were confirmed by inoculation of soybean seedlings with the isolates. Characteristics of the reisolated endosymbiont from induced root nodules were identical to those of the first isolate. From these results, it was confirmed that Bradyrhizobium strain isolated from the root nodules of Glycine max was a real symbiont, and was named B. japonicum SNU001.

Bradyrhizobium sp. SNU001 nod 유전자 클로닝

고세리,심웅섭,안정선 한국미생물학회 1992 미생물학회지 Vol.30 No.4

대두 (Glycine max) 뿌리혹의 질소고정 공생균주 Bradyrhizobium japonicum SNU001 의 nod 유전자를 클로닝하였다. Rhizobium meliloti 의 4.5 kb EcoRI/ HindIII 절편을 탐침으로 한 게놈 혼성화 반응을 분리균주의 게놈상에 nod 유전자가 존재함을 확인하고, lambda EMBL3-BamHI vector 를 이용하여 genomic library 를 작성하였다. 작성된 library 로부터 1, 2 차 선별과정을 통해 nod 유전자가 있는 클론 1-5 를 선별하고, 클론 2로부터 nod DABC 탐침과 lambda DNA 탐침을 사용한 혼성화반응을 수행하여 삽입된 genomic DNA 에 대한 부분적인 제한효소 지도를 작성하였다. nod DABC 탐침과 가장 강한 혼성화반응을 보인 phage 클론 lambda CNS-1 의 3.9 kb BamHI 절편을 pBS KS(+) vector 에 subcloning 하고 동일한 탐침을 이용한 혼성화반응을 통해 subclone pBjCNS-1 을 선별하였다. 이 subclone 에 대한 부분적인 제한효소 지도를 작성하여 nod DABC 가 1.8 KpnI/SacI 절편에 존재함을 확인하였다. Molccular cloning of nod genes from Bradvrhizobium sp. SNU001, a nitrogen-fixing symbiont isolated from thc root nodules of soybean (Clycine trim) . was carried out. nod genes were found to be located on thc genome of the symbiont by gcnomic hybridization with 4.5 kb EcoRI/HndIII fragment (nod DABC) of Rhizohium meliloti as probe. Genomic library of this symbiont was constructed using h phage EMBL3-BanlHI vector. from which five nod positive clones were sclectcd by primary and secondary screening methods. The partial restriction map of inserted genomic DNA of h CNS-l(c1one 2) was constructed. and 3.9 kh Bun7HI fragment. which showed strong hybridization signal to the probe, was subcloned into pBS KS(+) plasmid vector. Partial restriction inap ot' a selected subclone (pBjCNS-I) was constructed and nod DABC was found to be located on the 1.8 kb KpnI/Sacl fragment of this subclone.

유전자교정 기반 신육종기술의 글로벌 특허동향

고세리(Serry Koh),최유리(Youri Choi),이주영(Joo Young Lee),장지영(Jiyoung Jang),최규환(Kyuwhan Choi) 한국육종학회 2021 한국육종학회지 Vol.53 No.4

New Breeding Technology (NBT) refers to gene editing technologies that are used to develop crop plants with beneficial traits,from biotic/abiotic resistance to nutritional enhancement, including zinc finger nucleases (ZFN), transcription activator-like effector nucleases,clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9, meganucleases, and oligo directed mutagenesis. A total of 1,119 valid NBT patents were analyzed in this study to examine global trends in the patent and market expansion strategies formajor patent applicants. Based on the claims specified, valid patents in each patent office were analyzed through the applicant’s country oforigin, field of technology, and plant/crop species. Patents claiming applications of CRISPR-based technology to major crop plants, includingrice, corn, wheat, tomato, and canola, have rapidly increased in the China National Intellectual Property Administration (CNIPA) since 2013. The patent family size (PFS) can be used as an indicator of intellectual property (IP)-based market expansion strategies and target marketsof interests of patent applicants. Many university- and research-oriented institution Chinese applicants showed low PFS (2.1) because theyfiled patents mostly in CNIPA. In contrast, high PFS of US and German (DE) applicants such as Corteva Agriscience (US), KWS SAATAG (DE), Cellectics (FR), and Syngenta Participations AG (CH) represented their active strategies for global gene-edited crop market expansion. Corteva Agriscience (US, 238 patents) was the global leader in patents using NBT, ranging from ZFN to CRISPR-based technologies appliedto most major crops, including corn, soybean, and wheat.

Arabidopsis TTR1 Causes LRR-Dependent Lethal Systemic Necrosis, rather than Systemic Acquired Resistance, to Tobacco Ringspot Virus

남문,고세리,김성욱,Leslie L. Domier,전재흥,Su-Heon Lee,Andrew F. Bent,문제선 한국분자세포생물학회 2011 Molecules and cells Vol.32 No.5

Most Arabidopsis ecotypes display tolerance to the To-bacco ringspot virus (TRSV), but a subset of Arabidopsis ecotypes, including Estland (Est), develop lethal systemic necrosis (LSN), which differs from the localized hypersensitive responses (HRs) or systemic acquired resistance (SAR) characteristic of incompatible reactions. Neither viral replication nor the systemic movement of TRSV was restricted in tolerant or sensitive Arabidopsis ecotypes; therefore, the LSN phenotype shown in the sensitive ecotypes might not be due to viral accumulation. In the present study, we identified the Est TTR1 gene (tolerance to Tobacco ringspot virus 1) encoding a TIR-NBS-LRR protein that controls the ecotype-dependent tolerant/sensitive phenotypes by a map-based cloning method. The tolerant Col-0 ecotype Arabidopsis transformed with the sensitive Est TTR1 allele developed an LSN phenotype upon TRSV infection, suggesting that the Est TTR1 allele is dominant over the tolerant ttr1 allele of Col-0. Multiple sequence alignments of 10 tolerant ecotypes from those of eight sensitive ecotypes showed that 10 LRR amino acid polymorphisms were consistently distributed across the TTR1/ttr1 alleles. Site-directed mutagenesis of these amino acids in the LRR region revealed that two sites, L956S and K1124Q, completely abolished the LSN phenotype. VIGS study revealed that TTR1 is dependent on SGT1, rather than EDS1. The LSN phenotype by TTR1 was shown to be transferred to Nicotiana benthamiana, demonstrating functional conservation of TTR1 across plant families, which are involved in SGT-dependent de-fense responses, rather than EDS1-dependent signaling pathways.

The Crystal Structure of L-Leucine Dehydrogenase from Pseudomonas aeruginosa

Seheon Kim,고세리,Wonchull Kang,Jin Kuk Yang 한국분자세포생물학회 2022 Molecules and cells Vol.45 No.7

Leucine dehydrogenase (LDH, EC 1.4.1.9) catalyzes the reversible deamination of branched-chain L-amino acids to their corresponding keto acids using NAD+ as a cofactor. LDH generally adopts an octameric structure with D4 symmetry, generating a molecular mass of approximately 400 kDa. Here, the crystal structure of the LDH from Pseudomonas aeruginosa (Pa-LDH) was determined at 2.5 Å resolution. Interestingly, the crystal structure shows that the enzyme exists as a dimer with C2 symmetry in a crystal lattice. The dimeric structure was also observed in solution using multiangle light scattering coupled with size-exclusion chromatography. The enzyme assay revealed that the specific activity was maximal at 60°C and pH 8.5. The kinetic parameters for three different amino acid and the cofactor (NAD+) were determined. The crystal structure represents that the subunit has more compact structure than homologs’ structure. In addition, the crystal structure along with sequence alignments indicates a set of non-conserved arginine residues which are important in stability. Subsequent mutation analysis for those residues revealed that the enzyme activity reduced to one third of the wild type. These results provide structural and biochemical insights for its future studies on its application for industrial purposes.

고추의 게놈 분석

안정선(Chung Sun An),김성천(Sung Cheon Kim),고세리(Se Li Go) 한국식물학회 1996 Journal of Plant Biology Vol.39 No.1

The genome of red pepper was investigated by thermal denaturation, reassociation kinetics and measurement of nuclear volume for its base composition, spectrum of kinetic components and genome size. Base composition was estimated to be 37% (G+C) based on melting temperature. The reassociation of 300 nt fragments analyzed by hydroxyapatite chromatography revealed the presence of three kinetic components differing in fraction of genome, kinetic complexity and number of copies as follows; 4.8% (fast) with 5.6×10 exp (4) bp and 10,754, 26% (intermediate) with 1.9×10 exp (6) bp and 177, and 65% (slow) with 8.48×10 exp (8) bp and 1. These measurements demonstrate that the genome of red pepper has a 1C DNA content of 1.25×10 exp (9) bp, which is about 33% of 4.05×10 exp (9) bp calculated from nuclear volume of 62.4 μ㎥/IC.

An Expressed Sequence Tag Analysis for the Fast-Growing Shoots of Bambusa edulis Murno

조은정,Yurry Um,Seung Kwan Yoo,이형석,김호방,고세리,Hyun Cheol Shin,Yi Lee 한국식물학회 2011 Journal of Plant Biology Vol.54 No.6

Bamboo is one of the fastest growing plants in the world and is an economically important crop species in Asia. To identify the genes involved in fast shoot growth, an expressed sequence tag analysis was performed on Bambusa edulis Murno fast-growing shoots. Sequencing of the cDNA clones generated 1,402 5′-end high-quality expressed sequence tags (JG296384-JG297785, average length 655 bp),of which 1,101 clusters (143 consensus and 958 singletons)were revealed by sequence comparison to be unique and 597(54% of total clusters) of them have a putative ATG start codon. A Basic Local Alignment Search Tool X analysis showed that 995 of these genes were similar to genes present in the National Center for Biotechnology database. A total of 868 genes were most similar to rice genes. The most abundant genes were three thionin-coding genes, which have 27, 17, or 10 clones, respectively, followed by aminocyclopropanecarboxylate oxidase and cysteine protease. Thionin and putative cell elongation-associated genes, xyloglucan endotransglycosylase/hydrolase, expansin, cellulose synthase, and pectin esterase were analyzed by real-time reverse transcription polymerase chain reaction using genespecific primers. These results suggest that this high-quality library could be a good resource for understanding molecular events of bamboo shoot elongation, and the full-length clones could be used for crop improvement studies in the future.