Earticle

SS10 microbial strain, which is isolated from soil of sweet sorghum field, was characterized and identified as Staphylococcus sp. by analysis of 16S rDNA sequence and biochemical studies, and named as Staphylococcus sp. SS10 which has high xylanase activities. The optimum temperature and pH for xylanase activity of Staphylococcus sp. SS10 were 50℃ and 10.0, respectively. The xylanase activity was strongly inhibited by Al+++. The xylanase gene was cloned from Staphylococcus sp. SS10 genomic DNA by polymerase chain reaction (PCR). The amplified PCR product was ligated with the T&A cloning vector system and the constructed plasmids were transformed into E. coli DH5α. The sequence analysis of the insert DNAs revealed the identification of a 640-bp region containing xylanase open reading frame. According to xylanase gene sequence analysis, Staphylococcus sp. SS10 had gene sequence similarity of 99% with Bacillus subtilis Xyl gene for xylanase (AB457186.1).

질병을 야기하는 병원성균주들의 특성 및 이의 선택적 치료는 생명공학의 가장 핵심적인 연구분야 중의 하나이 다. 페니실린의 발견 이후 지속적인 연구를 통하여 병원성 균주를 선택적으로 제거할 수 있는 항생제들이 성공적으 로 개발되어져 왔으나, 최근에는 항생제 내성균주들의 출 현으로 한계에 부딪히고 있는 실정이다. 따라서 병원성균 주 혹은 질병세포를 선택적으로 인식․공격․제거 할 수 있 는 새로운 질병치료 패러다임이 필요한 실정이다. 식중독 을 일으키는 대표적인 미생물인 살모넬라균의 Type III Secretion System (T3SS)는 동물세포에 직접 단백질을 주 입(injection)할 수 있으며(1), 이를 활용한 라이브백신의 모델 시스템으로서 많은 각광을 받고 있다(Fig. 1). T3SS는 많은 그람음성균주들과 동물 혹은 식물 숙주 (host)들과의 interaction에 핵심적인 기관이며, 박테리아의 운동성을 담당하는 편모로부터 진화되어졌다고 알려져 있 다(1, 2). T3SS의 기본구조는 편모와 매우 유사하며, T3SS의 바늘 구조물의 지름은 8 nm이며 길이는 80 nm이다(3, 4). 살모넬라균이 산소농도, 삼투압 등의 외부조건을 인식하여 소장에 도달하였음을 인식하면, Salmonella Pathogenicity Island-1 (SPI-1) network을 활성화하여 T3SS 구조물 단 백질 들을 합성하여 균주외부에 발현시킨다(Fig. 1). 이와 동시에 effector라는 일련의 단백질들을 발현한 후, T3SS 를 통하여 숙주인 소장세포 내부로 주입 (injection)한다. 주입된 effector들은 숙주세포의 신호전달네트워크 (signal transduction network)를 조작하여 숙주세포의 변형을 유 발하고, 변형된 인체 소장세포 내부로 살모넬라균이 침입 한 이후 혈관을 통하여 인체의 다른 기관으로 이동하여 식중독 등의 질병을 일으킨다. 살모넬라균이 인체침투 및 질병유발을 위해 사용하는 T3SS를 이용하면, 암 및 질병세포 내부에 선택적으로 치 료용 단백질을 직접적으로 주입하는 것이 가능하므로, 암 세포 치료 및 질병치료의 새로운 패러다임으로 많은 기대 를 모으고 있다. 이를 위하여 특정 세포나 주변조건을 인 식하여 목적 유전자를 발현시키는 artificial gene circuit을 만드는 연구도 활발히 이루어지고 있다(5, 6). 본 연구진 은 T3SS를 이용한 외래단백질 분비 (secretion) 가능성을 살펴보기 위하여, 살모넬라균의 effector 단백질과 모델단 백질인 lipase와의 fusion protein을 제작하고 이를 균주외 부로 분비하는 연구를 수행하였다.

New protein secretion system was developed using Type III Secretion System of Salmonella. N-terminal region of SlrP and SptP effector proteins were fused with TliA and EstA-P lipases by overlapping PCR. Lipase activity of Salmonella with SptP-TliA fusion system increased by 2.6 fold compare with wild type Salmonella strain. This result showed that lipase secretion via the T3SS would be a useful protein secretion machinery.

Targeting particular mRNAs has become an excellent approach for the silencing of gene expression. Here, we showed a cell-penetrating antibody (transbody) that specifically hydrolyzes targeted mRNAs in the cellular cytosol and leads to targeted gene silencing. We generated a synthetic library on the yeast surface based on 3D8 VL, which is a cell-penetrating and nucleic acid-hydrolyzing single domain antibody. With the cell-penetrating activity, 3D8 VL into the cytosol of living cells can selectively decrease the amount of target sequence-carrying mRNAs. We selected 3D8 VL variants had higher affinity and greater selective hydrolyzing activity for target ss-DNAs than for off targets in the library by using 18-bp single-stranded (ss)-DNAs as target substrates. In particular, one 3D8 VL variant targeting the Her2 sequence showed more efficient downregulation of Her2 expression than a small-interfering RNA targeting the same Her2 sequence, and the variant caused apoptotic cell death of Her2-overexpressing breast cancer cells. Our results demonstrate that the 3D8 VL variants with cell penetrating, nucleic acid-hydrolyzing activity and sequence-selectivity could degrade target mRNAs in the cytosol, which suggests that they would be potential toolds in anti-cancer, anti-viral therapies.

Gene silencing by targeting specific genes for degradation, particularly at the mRNA level, is an invaluable tool for gene function analysis and a powerful therapeutic strategy for human diseases, including cancer and viral infections. For degrading cytosolic RNAs is the use of protein-based RNases and DNA/RNA-hydrolyzing mAbs, which can penetrate into living cells and degrade cytosolic RNAs. However, these approaches non sequence–specificity, leading to significant cytotoxicity. Based on a cell-penetrating, nucleic acid-hydrolyzing, 3D8 VL[1], we generated a synthetic library on the yeast surface by randomizing potential base interacting residues located in surface of c,c’,f-strands. And we considered that selected randomizing residues in framework by fixing several framework residues that affect directly or indirectly to CDR and antibody stability, like upper core and lower core, charged cluster. We isolated 3D8 VL variants with classical swine fever virus Npro and E2 genes –selective binding against 18-bp single stranded (ss)-RNA substrates, selected 3D8 VL variants that had ~10-fold higher affinity and ~2-fold greater selective affinity for target ss-RNAs than for off targets.

Targeting particular mRNAs has become an excellent approach for gene silencing. Here, we showed a cell-penetrating antibody (transbody) that specifically hydrolyzes targeted mRNAs in the cellular cytosol and leads to targeted gene silencing. We generated a synthetic library on the yeast surface based on 3D8 VL, which is a cell-penetrating and nucleic acid-hydrolyzing single domain antibody of the light chain variable domain. With the cell-penetrating activity, 3D8 VL into he cytosol of living cells can selectively decrease the amount of target sequence-carrying mRNAs. We selected 3D8 VL variants had higher affinity and greater selective hydrolyzing activity for target single-stranded (ss)-DNAs than for off targets in the library by using 18bp-ssDNAs as target substrates. In particular, one 3D8 VL variant targeting the Her2 sequence showed more efficient downregulation of Her2 expression than a smallinterfering RNA targeting the same sequence, and the variant caused apoptotic cell death of Her2-overexpressing breast cancer cells. Our results demonstrate that the 3D8 VL variants with cell penetrating, nucleic acid-hydrolyzing activity and sequence-selectivity could degrade target mRNAs in the cytosol, which suggests that they would be potential tools in anti-cancer or anti-viral therapies.

Gene silencing by targeting specific genes for degradation, particularly at the mRNA level, is an invaluable tool for gene function analysis and a powerful therapeutic strategy for human diseases, including cancer and viral infections. For degrading cytosolic RNAs is the use of protein-based RNases and DNA/RNAhydrolyzing mAbs, which can penetrate into living cells and degrade cytosolic RNAs. However, these approaches non sequence–specificity, leading to significant cytotoxicity. Based on a cell-penetrating, nucleic acid-hydrolyzing mAb, 3D8 VL, we generated a synthetic library on the yeast surface by randomizing potential base interacting residues located in surface of C, C’, F-strand, near F-strand framework 103, 105 residue. And we considered that selected randomizing residues in framework by fixing several framework residues that affect directly or indirectly to CDR and antibody stability, like upper core and lower core, charged cluster. Classical swine fever (CSF) or hog cholera (also sometimes called pig plague) is a highly contagious disease of pigs and wild boar. The infectious agent responsible is a virus CSFV. We isolated 3D8 VL variants with classical swine fever virus Npro and E2 genes–selective binding against 18-bp single stranded (ss)DNA substrates that substitute deoxy-uridin for thymine, selected 3D8 VL variants.

Chemical synthesis of 1,2-propanediol (1,2-PD, propylene glycol) that is most commodity chemicals have some problems [1]. In this work, microbial fermentation was used form the production of useful industrial chemicals [2]. Saccharomyces cerevisiae was selected a host strain. Methylglyoxal synthase (mgs) and glycerol dehydrogenase (gldA), the two enzymes were selected for the production of 1,2-PD metabolism. And the two enzymes that are essential intermediate from DHAP and glycerol dehydrogenase [2-4]. The yeast expression vector, pESC-URA has multi-cloning sites, was used construction plasmid containing target gene mgs and gldA. The plasmid containing mgs and gldA was inserted into S. cerevisiae by LiAc/SS-carrier DNA/PEG transformation. And then strain containing plasmid was screened by SDC (A,T) medium. The culture medium was YPD adding galactose as trigger for the activation of GAL promoter in the plasmid [5]. The concentration of 1,2-PD was analyzed by HPLC with refractive index detector. The mobile phase was 0.5M sulfuric acid and flow rate was 0.5 ml/min. After growing the yeast, we broke the cells to extract 1, 2-PD. The solution for 1,2-PD analysis was condensed and analyzed by HPLC. Cells containing mgs or gldA or two genes in pESC-URA vector were shown the production of 1,2-PD.

Sequence similarity in biological databases is used to characterize a newly discovered protein and confirming the existence of its homologs. This is often computationally very expensive. We have implemented a new algorithm that performs sequence similarity search using a pre-search phase. The proposed algorithm works in three phases. As a pre-preparation for Pre-Search, we locate a sequence, similar to the query sequence to extract all common words between the former and the latter. In the second phase, the pre-search phase, we locate all sequenes containing any of the randomly chosen common words. The list is further scanned in the third phase and the results obtained from the second phase are refined using Similarity Search (SS) algorithm, described in the paper. We have preprocessed the Uniref100.FASTA protein database containing 9,757,328 records downloaded from uniprot.org, to suit our application of sequence similarity search. The algorithm is simple and can be applied in various perspectives. These include searching in DNA and protein sequence databases, motif finding, and gene identification search. Pre-Search reduces the search space using much faster simpler algorithm. In large database search, its effect could be phenomenal.

열충격 시그마인자를 코딩하는 유전자 rpoH가 결여된 돌연변이체 대장균(Escherichia coli satrain A7448)을, 메탄올 자화세균인 Methylovorus sp. strain SS1 DSM11726의 phagemid library로 형질전환 시켜서 $30^{\circ}C$에서 성장하는 Escherichia coli strain A7448 로부터 Methylovorus sp. strain SS1 DSM11726의 rpoH 유전자를 클로닝하고 그 염기서열을 분석하였다. 1,793-bp 염기서열 분석 결과 Methylovorus sp. strain SS1 DSM11726의 RpoH는 284개의 아미노산으로 이루어져 있었으며 예상된 분자량은 32,006, p1값은 5.79로 나타났으며, 동일계열의 ${\beta}$-proteobacteria에 속하는 세균들의 RpoH와 높은 상동성을 보여주었다. Methylovorus sp. strain SS1 DSM11726의 RpoH는 대장균의 RpoH의 기능을 대신할 수 있음을 보여주었다. 열충격 후 RpoH양은 15분까지 지속적으로 증가하다 20분 뒤 양이 감소하는 양상을 나타내었다. 이는 Methylovorus sp. strain SS1 DSM11726의 RpoH 단백질 역시 열에 의해 유도됨을 말해 준다.

Using complementation of RpoH deficient E. coli strain A7448, the rpoH gene encoding heat shock sigma factor 32 (${\sigma}^{32}$) from Methylovorus sp. strain SS1 DSM11726 was cloned and sequenced. Sequence analysis of a stretch of 1,796-bp revealed existence of an open reading frame encoding a polypeptide of 284 amino acid (32,006 dalton). Deduced amino acid sequence of the Methylovorus sp. strain SS1 RpoH showed that 59.6%, 39.1% and 51.4% identities with those of Nitrosomonas europaea (${\beta}$-proteobacteria), Agrobacterium tumefaciens ($\alpha$-proteobacteria) and E. coli (${\gamma}$-proteobacteria). The expression level of the functional ortholog of RpoH of Methylovorus sp. strain SS1 was increased transiently after heat induction, further indicating that it functions as a heat shock sigma factor.

Sequence analysis of the Chlorella virus SS-2 revealed one putative nuclease gene that is 807 bp long and encodes a 31kDa protein. Multiple sequence alignment analysis reveals the presence of highly conserved PD-(D/E)XK residues in the encoded protein. The gene cloned into an expression vector was expressed as a His-tagged fusion protein in chaperone containing pKJE7 cells. The recombinant protein was purified using a His-Trap chelating HP column and used for functional analysis. Exonuclease activity of the SS-2 nuclease was detected when the DNA substrates, such as linear ssDNA, PCR amplicon, linear dsDNA with 5'-overhang ends, 3'-overhang ends, or blunt ends were used. Covalently closed circular DNA was also degraded by the SS-2 recombinant protein, suggesting that the SS-2 nuclease has an endonuclease activity. Stable activity of SS-2 nuclease was observed between $10^{\circ}C$ and $50^{\circ}C$. The optimum pH concentrations for the SS-2 nuclease were pH 6.0-8.5. Divalent ions inhibited the SS-2 nuclease activity.

The welding fume has been implicated as a causal agent in respiratory disease such as pneumoconiosis. The molecular mechanism by which welding fume induces toxicity in the lung is still unknown, but studies have focused on histological structure and indirect approach measuring the pulmonary damage markers. In the present study, gene expression profiles were analyzed in the lung of rats exposed by manual metal-arc stainless-steel (MMA-SS) welding fume for 30 days using Affymetrix GeneChip$^{(R)}$. Totally, 379 genes were identified as being either up- or down-regulated over 2-fold changes (P<0.01) in the lung of low- or high-dose group and were analyzed by using hierarchical clustering. We focused on genes involved in immune/inflammation responses were differentially regulated during lung injury induced by welding fume exposure. The information of these deregulated genes may contribute in elucidation of the inflammation mechanism during lung injury such as lung fibrosis.

Hypernodulation soybean mutant, SS2-2, is characterized with greater nodulation and nitrogen fixing ability in the root nodule than its wild type, Shinpaldalkong 2. The present study was performed to identify a genetic locus conferring hypernodulation in soybean mutant SS2-2 and to determine whether the gene controlling the hypernodulation of SS2-2 is allelic to that controlling the supernodulation of nts382 mutant. Hybridization studies between SS2-2 and Taekwangkong revealed that the recessive gene was responsible for the hypernodulation character in soybean mutant SS2-2. Allelism was also tested by crossing supernodulating mutant nts382 and hypernodulating mutant SS2-2 that both hypernodulation and supernodulation genes were likely controlled by an identical locus. Molecular marker mapping of hypernodulation gene in SS2-2 using SSR markers confirmed that the gene conferring hypernodulation was located at the same loci with the gene conferring supernodulation. It is interesting to note that the same gene controlled the super- and hyper-nodulation characters, although SS2-2 and nts 382 exhibited differences in the amount of nodulation in the root system. Further genetic studies should be needed to clarify the genetic regulation of super- and hyper-nodulation in soybean.

The short petiole trait is valuable for the development of plant ideotype with high yield. Soybean breeding line, SS98206SP, showed extremely short petioles in greenhouse and field. In this study, the short petiole of two mutant lines, SS98206SP and D76-1609, were investigated to determine the genetic segregations. These two mutants were crossed with each other and with two normal petiole genotypes. Genetic analysis indicated that the short petioles in D76-1609 and SS98206SP were controlled by a single recessive gene, respectively. The short petiole gene in SS98206SP was non-allelic with lps, conferring short petiole in D76-1609. Two recessive genes showed complementary relationship having short petioles with recessive homozygote (LPS1-lps?lps?, lps1lps1LPS?-, lps1lps1lps?lps?). Our data indicated that the short petioles in SS98206SP were controlled by a single recessive gene designated as lps3.

Somatostatin (SS) and growth hormone-releasing hormone (GHRH) are primary factors regulating growth hormone (GH) secretion in the pituitary. To date, it remains unknown how this rhythm is controlled endogenously, although there must be coordination of circadian manners. Melatonin was the main regulator in biological rhythms, and its secretion has fluctuation by photic information. But relationship between melatonin and growth-related genes (ghrh and ss) is unclear. We investigated circadian rhythms of melatonin secretion, ghrh and ss expressions, and correlation between melatonin with growth-related genes in tiger puffer Takifugu rubripes. The melatonin secretion showed nocturnal rhythms under light and dark (LD) conditions. In constant light (LL) condition, melatonin secretion has similar patterns with LD conditions. ss1 mRNA was high during scotophase under LD conditions. But ss1 rhythms disappeared in LL conditions. Ghrh appeared opposite expression compared with melatonin levels or ss1 expression under LD and LL. In the results of the melatonin injection, ghrh and ss1 showed no significant expression compared with control groups. These results suggested that melatonin and growth-related genes have daily or circadian rhythms in the tiger puffer. Further, we need to know mechanisms of each ss and ghrh gene regulation.

Bacterial leaf pustule (BLP) caused by Xanthomonas axonopodis pv. glycines is a serious disease to make pustule and chlorotic haloes in soybean [Glycine max (L). Merr.]. While inheritance mode and map positions of the BLP resistance gene, rxp are known, no sequence information of the gene was reported. In this study, we made five near isogenic lines (NILs) from separate backcrosses (BCs) of BLP-susceptible Hwangkeumkong $\times$ BLP-resistant SS2-2 (HS) and BLP-susceptible Taekwangkong$\times$ SS2-2 (TS) through foreground and background selection based on the four-stage selection strategy. First, 15 BC individuals were selected through foreground selection using the simple sequence repeat (SSR) markers Satt486 and Satt372 flanking the rxp gene. Among them, 11 BC plants showed the BLP-resistant response. The HS and TS lines chosen in foreground selection were again screened by background selection using 118 and 90 SSR markers across all chromosomes, respectively. Eventually, five individuals showing greater than 90% recurrent parent genome content were selected in both HS and TS lines. These NILs will be a unique biological material to characterize the rxp gene.

Objective : The purpose of this study was to investigate the possible association of estrogen receptor alpha ($ER{\alpha}$) gene polymorphisms in a cohort of degenerative spondylolisthesis (DS) patients. Methods : Accordingly, the authors examined the association between DS and $ER{\alpha}$ gene polymorphisms in 174 patients diagnosed with DS. The $Pvu$ $II$ and $Xba$ $I$ polymorphisms, bone mineral density at the lumbar spine and femoral neck, and biochemical markers were analyzed and compared in the 174 patients with DS and 214 patients with spinal stenosis (SS). Results : A comparison of genotype frequencies in DS and SS patients revealed a significant difference for the $Pvu$ $II$ polymorphism only ($p$=0.0452). No significant difference was found between these two groups with respect to the $Xba$ $I$ polymorphism, BMD or biochemical markers. No significant association was found between the$Pvu$ $II$ polymorphism of $ER{\alpha}$ and BMD, vertebral slip or biochemical markers in patients with DS. Conclusion : These results suggest that the $ER{\alpha}$ gene polymorphism using $Pvu$ $II$ restriction enzyme influences the prevalence of DS.

The single nucleotide polymorphism (SNP) ss469415590 in the interferon lambda-4 (IFNL4) gene has recently been reported to have an association with treatment response in chronic hepatitis C. However, any importance of the SNP in association with response to pegylated interferon (PEG-IFN) therapy in patients with chronic hepatitis B (CHB) is unclear. We retrospectively analyzed data for Thai patients with CHB treated with PEG-IFN for 48 weeks. Virological response (VR) for HBeAg-positive CHB was defined as HBeAg seroconversion plus HBV DNA level <2,000 IU/mL at 24 weeks post-treatment. VR for HBeAg-negative CHB was defined as an HBV DNA level <2,000 IU/mL at 48 weeks. The SNP was identified by real time PCR using the TaqMan genotyping assay with MGB probes. A total 254 patients (107 HBeAg-positive and 147 HBeAg-negative) were enrolled in the study. The distribution of TT/TT, ${\Delta}G/TT$ and ${\Delta}G/{\Delta}G$ genotypes was 221 (87.0%), 32 (12.6%) and 1 (0.4%), respectively. Patients with non-TT/TT genotypes had significantly higher baseline HBV DNA levels than patients with the TT/TT genotype. In HBeAg-positive CHB, 41.2% of patients with TT/TT genotype versus 50.0% with non-TT/TT genotype achieved VR (P=0.593). In HBeAg-negative CHB, the corresponding figures were 40.3% and 43.5%, respectively (P=0.777). There was no significant correlation between the SNP genotypes and HBsAg clearance in both groups of patients. In summary, ss469415590 genotypes were not associated with response to PEG-IFN in Thai patients with HBeAg-positive and HBeAg-negative CHB.

Xanthomonas oryzae pv. oryzae (Xoo) produces a putative effector, XoAvrBs2. We expressed XoAvrBs2 homologously in Xoo with a TAP-tag at the C-terminus to enable quantitative analysis of protein expression and secretion. Addition of rice leaf extracts from both Xoo-sensitive and Xoo-resistant rice cultivars to the Xoo cells induced expression of the XoAvrBs2 gene at the transcriptional and translational levels, and also stimulated a remarkable amount of XoAvrBs2 secretion into the medium. In a T3SS-defective Xoo mutant strain, secretion of the TAPtagged XoAvrBs2 was blocked. Thus, we elucidated the transcriptional and translational expressions of the XoAvrBs2 gene in Xoo was induced in vitro by the interaction with rice and the induced secretion of XoAvrBs2 was T3SSdependent. It is the first report to measure the homologous expression and secretion of XoAvrBs2 in vitro by rice leaf extract. Our system for the quantitative analysis of effector protein expression and secretion could be generally used for the study of host-pathogen interactions.

Background: Several studies have previously focused on associations between the (GT)n repeat polymorphism of the heme oxygenase-1 (HO-1) gene promoter region and risk of cancers, but results are complex. We conducted the present meta-analysis to integrate relevant findings and evaluate the association between HO-1(GT)n repeat polymorphism and cancer susceptibility. Materials and Methods: Published literature was retrieved from the PubMed/MEDLINE, EMBASE and ISI Web of Science databases before November 2013. For all alleles and genogypes, odds ratios were pooled to assess the strength of the associations using either fixed-effects or random-effects models according to heterogeneity. Subgroup analysis was conducted according to ethnicity and histopathology. Results: A total of 10 studies involving 2,367 cases and 2,870 controls were identified. The results showed there was no association between HO-1 (GT)n repeat polymorphism and the cancer risk both at the allelic and genotypic level. However, in the stratified analysis, we observed an increased risk of squamous cell carcinoma in persons carrying the LL genotype and the LL+LS genotype as compared with those carrying the SS genotype. When the LS and SS genotypes were combined, the odds ratio for squamous cell carcinoma in LL-genotype carriers, were also significantly increased. No publication bias was observed. Conclusions: The LL genotype and L-allele carrying genotypes (LL+LS) of HO-1 (GT)n repeat polymorphism are potential genetic factors for developing squamous cell carcinoma. More large and well-designed studies are required for further validations.

Xanthomonas axonopodis pv. glycines(Xag) is a pathogen that causes bacterial leaf pustule(BLP) disease in soybeans grown in Korea and the southern United States. Typical and early symptoms of the disease are small, yellow to brown lesions with raised pustules that develop into large necrotic lesions leading to a substantial loss in yield due to premature defoliation. After Xag infects PI 96188, only pustules without chlorotic haloes were observed, indicating the different response to Xag. To identify differentially expressed genes prior to and 24 hr after Xag inoculation to PI 96188 and BLP-resistant SS2-2, an oligonucleotide macroarray was constructed with 100 genes related to disease resistance and metabolism from soybean and Arabidopsis. After cDNAs from each genotype were applied on the oligonucleotide macroarrays with three replicates and dye swapping, 36 and 81 genes were expressed as significantly different between 0 hr and 24 hr in PI 96188 and SS2-2, respectively. Six UniGenes, such as the leucine-rich repeat protein precursor or 14-3-3-like protein, were selected because they down-regulated in PI 96188 and up-regulated in SS2-2 after Xag infection, simultaneously. Using tubulin and cDNA of Jangyeobkong(BLP-susceptible) as controls, the oligonucleotide macroarray data concurred with quantitative real-time RT-PCR(QRT RT-PCR) results in most cases, supporting the accuracy of the oligonucleotide macroarray experiments. Also, QRT RT-PCR data suggested six candidate genes that might be involved in a necrotic response to Xag in PI 96188.