• BMB Reports
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• 제30권3호
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• pp.173-176
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• 1997

We isolated a cDNA clone that encodes a ribulose-1,5-bisphosphate carboxylase small subunit (rbcS) from spinach using a soybean rbcS cDNA as a probe. The small subunit consists of 180 amino acids including a transit peptide of 57 residues. Comparison of the amino acid sequence with those of other plant species shows a maximum of 70-80% identical residues. Southern blot analysis suggests the existence of multiple rbcS genes in the spinach genome. Northern blot analysis indicates that the rbcS gene is expressed predominantly in leaves and that the expression of the gene is induced by light.

• Journal of Photoscience
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• 제10권3호
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• pp.245-249
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• 2003

The psbA gene of photo system II was cloned and characterized from the P. ginseng chloroplast. The psbA gene is composed of 1,062 nucleotides. The overall amino acid sequence shows 99% and 98% identities to dicots and monocots of higher plants, respectively. Southern blot analysis revealed that a single copy of the psbA gene existed in the chloroplast genome. Northern blot analysis of the in vivo accumulation of the psbA transcript, after being grown under the different intensities (5%, 10%, 20%, and 100%) of daylight, indicated that the steady-state level of the psbA transcript was not significantly affected by light intensity.

• BMB Reports
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• 제39권3호
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• pp.263-269
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• 2006

In the genome of Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus, open reading frame 39 (Ha39) is the only gene predicted to encode an RNA recognition protein. Computer analysis revealed that Ha39 homologues were found in 15 NPVs, but not in GVs. Its transcripts were detected from 3 through 72 hours post infection (h p.i.) using RT-PCR and Northern blot analysis. The protein was detected in infected-cell lysates from 6 h p.i. Western blot assay of ODV and BV preparations revealed that Ha39 encodes a structural protein associated with BVs. Additionally, immunofluorescence microscopy demonstrated that the protein was present within cytoplasm in virus-infected cells, but not in the nuclear region.

• Journal of Microbiology and Biotechnology
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• 제5권5호
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• pp.264-268
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• 1995

Sulforhodamine B(SRB) assay was performed on the human lung carcinoma, A549 cell line to screen soil microorganisms for production of anti-cancer agent. Among 4, 265 microorganisms, 45 isolates were selected for their cytotoxicity and tested for their effects on the expression of c-myc by RNA slot blot and Northern blot analysis resulting in selection of No.2303 isolate. This No.2303 was identified as Streptomyces sp. by ISP classification and the chemotaxonomic analysis method. NO.2303 inhibited the expression of cmyc in Col0320 DM and A549 cell lines. The culture extract of No. 2303 also inhibited the progression of the cell cycle of Go in NIH 313 cells, implying that the extract also inhibited the expression of c-myc in NIH 313 cell.

• 한국연초학회지
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• 제19권2호
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• pp.117-123
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• 1997

Viral coat protein (CP) encoding cDNA with artificial start and stop codons was synthesized by reverse-transcriptase polymerase chain reaction (RT-PCR) from the Korean isolate of potato virus Y-vein nectrosis strain (pVY-VN). To make PVY CP cDNA to untranslatable form, three stop codons were inserted near the start codon by "megaprimer-PCR" method. The untranslatable CP cDNA was subcloned to plant expression vector and transferred to N. tabacum cv. NC82 by Agrobacterium-mediated transformation. Highly resistant plants to PVY infection were screened, based on symptom development after mechanical virus inoculation. By genomic PCR and Southern blot analysis, one or more copies of the untranslatable CP gene were found in all transformants. From northern blot analysis, highly resistant transgenic lines had very low level of CP transcript but susceptible lines had high level, suggesting resistance to PVY infection should be related to RNA-mediated mechanism.mechanism.

• 한국환경성돌연변이발암원학회지
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• 제19권1호
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• pp.28-33
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• 1999

The SNF2 family includes proteins from a variety of species with roles I cellular processes such as transcriptional regulation, recombination and various types of DNA repair. Several proteins with unknown function are also included in this family. Here, we report the cloning and characterization of hrp 2+ gene (helicase related gene from S. pombe) which was isolated by PCR amplication using the conserved domain of SNF2 motifs within the ERCC6 gene which encodes a protein involved in DNA excision repair. The hrp2+ gene was isolated by screening with yeast S. pombe genomic library. The isolated cloned contained 6.5 kb insert DNA. Southern blot analysis confirmed that S. pombe chromosome contains the same DNA as hrp2+ gene and this gene exists as a single copy in S. pombe genome. The 4.7 kb transcript of mRNA was identified by Northern blot. To examined the transcriptional regulation of hrp2+ gene, DNA damaging agents were treated. These results indicated that the hrp2+ gene may not be directly involved in DNA replication, but may be involved in damage response pathway.

• Journal of Plant Biology
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• 제38권2호
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• pp.137-141
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• 1995

We isolated a cDNA clone, BLSC1, encoding 5' exon of ATP synthase CF0 subunit I from broccoli. BLSC1 is 285 nucleotides long which consists of a 5' noncoding region of 34 nucleotides, a 5' exon of 145 nucleotides and an intron of 106 nucleotides. The 5' exon codes for 48 amino acids which reveals mostly hydrophobic. The amino acid sequence deduced from BLSC1 shares 83%, 83% and 91% identities with the genes coding for atpF from wheat, rice and spinach, respectively. Genomic Southern blot analysis for BLSC1 showed a typically strong signal for a gene located in the chloroplast genome. Northern blot analysis identified three major classes of transcripts showing strong positive signals in the leaves, but only trace amounts of the transcripts were identified in the other organs like stems, flowr buds and roots.

• 식물조직배양학회지
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• 제25권2호
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• pp.131-134
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• 1998

Antisense PG 유전자 형질전판 토마토로부터 자식시켜 얻은 종자를 kanamycin 내성을 이용하여 5세대까지 분리, 육성하여 antisense PG 유전자가 안정적으로 고정된 식물체를 얻었다. 이들 식물체에는 genomic DNA gel blot 분석으로 antisense PG 유전자가 안정적으로 유전되며 northern blot 분석을 통하여 antisense RNA가 발현됨을 확인하였고, 또한 antisense RNA에 의한 endogenous PG 유전자의 발현 저해를 분석하였다. Antisense PG 유전자 형질전환 성숙 토마토내 PG 효소 활성이 비형질전환 성숙 토마토에 비하여 37-65% 수준으로 저하되었다.

• 원예과학기술지
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• 제32권1호
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• pp.91-99
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• 2014

본 연구는 배추에서의 저온 저항성 유전자를 개발하는데 목적이 있으며 이를 위해 먼저 저온($4^{\circ}C$) 스트레스가 처리된 내혼계배추를 대상으로 한 KBGP-24K oligo chip의 결과 [BrEMD(Brassica rapa EST and Microarray Database)]를 분석하였다. 그 결과 23,929개의 배추 unigene 중 저온 처리시 대조군 대비 5배 이상 발현이 증가하는 417개(1.7%)의 저온 반응 유전자를 1차 선발하고, 이들 중 기능이 정확히 알려지지 않았으나 완전장을 갖추고 있는 BrCSR로 명명한 유전자를 선발하였다. 이 유전자의 저온 저항성을 분석하기 위하여 형질전환용 과발현 vector인 pSL101 binary vector를 제작하여 담배에 형질전환시켰다. BrCSR이 과발현된 $T_1$ 세대 담배 형질전환체들은 PCR과 Southern hybridization 분석에 의해 선발하였고, BrCSR의 기능은 저온 처리 시 유전자의 발현 수준 분석과 표현형 검정을 통해 확인하였다. Quantitative real-time RT-PCR과 Northern blot hybridization 분석 결과, 형질전환 담배에서 BrCSR의 발현이 대조군보다 약 2배 정도 높게 발현되었으며 실제로 $4^{\circ}C$ 처리 후 표현형 분석에서 BrCSR이 과발현된 형질전환체들이 대조군보다 우수한 저온 저항성을 보여 주었다. 위 결과들에 근거하여 BrCSR 유전자가 저온 환경 하에서 식물의 생장과 저항성 향상에 중요한 역할을 담당하고 있음을 확인할 수 있었다.

• Applied Biological Chemistry
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• 제38권1호
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• pp.49-54
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• 1995

한국 마늘에 감염된 바이러스의 종류와 병 발생 메카니즘을 구명하기 위하여, 마늘 바이러스 cDNA clone들을 분리하였다. 24개 cDNA clone들의 부분적인 염기 서열을 결정하였고, 이 중 poly(A) tail을 가진 5개 clone들의 염기 서열을 결정하였다. 이를 이미 알려진 다른 식물 바이러스와 비교했을 때, clone V9은 일차구조가 carlavirus와 유사성을 보이므로 GLV cDNA clone으로 여겨진다. Northern blot 결과로부터 GLV genome의 크기는 8.5 knt이고, poly(A) tail을 가지고 있다는 것을 알 수 있었다. clone V9의 3' 말단부분에는 바이러스 복제과정에서 cis-acting element로 작용한다고 여겨지는 hexanucleotide motif(5'-ACCUAA)가 존재한다. 또한 carlavirus의 껍질 단백질 subgenomic RNA의 5' 말단에 보존되어 있는 5'-TTAGGT도 나타난다. 이들은 모두 carlavirus의 특징들이다. 껍질 단백질 유전자를 pRSET-A 발현 벡터에 재조합하고, E. coli BL21에서 발현시켰다. 발현된 껍질 단백질을 $Ni^{2+}$ NTA affinity chromatography에 의해 정제하였다. 껍질 단백질을 토끼에 주사하여 항체를 만든 후, immunoblot을 한 결과 GLV 껍질 단백질에 해당하는 24 kDa polypeptide가 인지되었다. 또한 다양한 마늘 품종에 대해서 immunoblot을 한 결과, GLV 껍질 단백질의 크기와 GLV의 감염정도가 마늘 품종에 따라서 차이가 있다는 것을 알 수 있었다.