• 제목/요약/키워드: full-length cDNA

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소 바이러스성 설사병 바이러스의 유전자 재조합 DNA clone의 작성에 관한 연구 (Construction of recombinant DNA clone for bovine viral diarrhea virus)

  • 여상건
    • 대한수의학회지
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    • 제32권3호
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    • pp.389-398
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    • 1992
  • Molecular cloning was carried out on the Danish strain of bovine viral diarrhea virus(BVDV) to construct strategy for the diagnostic tools and effective vaccine of BVD afterwards. A recombinant DNA clone(No. 29) was established successfully from cDNA for viral RNA tailed with adenine homopolymer at 3'-end. $^{32}P$-labeled DNA probes of 300~1,800bp fragments, originating from the clone 29, directed specific DNA-RNA hybridization results with BVDV RNA. Recombinant DNA of the clone 29 was about 5,200bp representing 41.6% of the full length of Danish strain's RNA, and restriction sites were recognized for EcoR I, Sst I, Hin d III and Pst I restriction enzymes in the DNA fragment.

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Molecular Cloning and Characterization of a Flavanone-3-hydroxylase Gene from Rubus occidentalis L.

  • Lee, Seung Sik;Lee, Eun Mi;An, Byung Chull;Barampuram, Shyamkumar;Kim, Jae-Sung;Cho, Jae-Young;Lee, In-Chul;Chung, Byung Yeoup
    • 방사선산업학회지
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    • 제2권3호
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    • pp.121-128
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    • 2008
  • Flavanone-3-hydroxylase (F3H) is one of the key enzymes for the biosynthesis of flavonals, anthocyanins, catechins and proanthocyanins. F3H catalyzes the $3{\beta}$-hydroxylation of (2S)-flavonones to form (2R, 3R)-dihydroflavonols. In this report, we isolated a full-length cDNA of RocF3H from black raspberry (Rubus occidentalis L.) using a reverse transcriptase-PCR and rapid amplification of the cDNA ends (RACE)-PCR. The full-length cDNA of RocF3H contains a 1,098 bp open reading frame (ORF) encoding a 365 amino acid protein with a calculated molecular weight of about 41.1 kDa and isoelectric point (pI) of 5.45. The genomic DNA analysis revealed that the RocF3H gene had three exons and two introns. Comparison of the deduced amino acid sequence of the RocF3H with other F3Hs revealed that the protein is highly homologous with various plant species. The conserved amino acids ligating the ferrous iron and the residues participating in the 2-oxoglutarate binding (R-X-S) were found in RocF3H at the similar positions to other F3Hs. Southern blot analysis indicated that RocF3H exist a multi-gene family. The isolation of RocF3H gene will be helpful to further study the role of F3H gene in the biosynthesis of flavonoids in R. occidnetalis.

인삼의 유용유전자원 확보를 위한 기능 유전체연구 (Functional Genomics for Mass Analysis of Useful Genes in Panax ginseng C.A. Meyer)

  • 양덕춘
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 2004년도 춘계 총회 및 학술대회
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    • pp.17-28
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    • 2004
  • 현재 재배되고 있는 고려인삼은 혼계상태로서 개체간의 형질변이가 대단히 심하며, 종자채취를 4년 후에 하기 때문에 신품종육성을 위해서는 매우 오랜 기간이 필요하다. 그러나 식물형질전환기술의 발달로 목적하는 유전자를 식물체에 재도입하여 새로운 품종을 육성할 수 있는 기술이 보급되어 유용유전자만 있다면 단시간에 고기능성 신품종을 육성할 수 있을 것이다. 본 과제에서는 인삼의 유용유전자를 대량으로 발굴하기 위하여 인삼의 조직별, 종간 및 처리간에 의한 cDNA library 총 10종 이상을 제작하고 이들 cDNA library로부터 인삼의 유용유전자원을 확보하기 위하여 EST 20,000개를 5' 한쪽 방향에서 분석하고 이들 분석된 EST clone의 데이터는 data base화 하여 많은 연구자들이 연구정보를 공유한 수 있게 하였다. 그리고 EST clone 중에서 완전한 단백질의 유전정보를 포함하고 있는 clone을100개 선발하여 전장의 염기서열(full length sequence)을 분석하고 data base를 구축하고parental clone을 선발하여 cDNA chip을 제작하였다. 특히 257 clone 주에서 기능성 및 내재해성 유용유전자를 선발하여 전염기서열분석(full length sequencing)을 한 후 인상에 재도입하여 고기능성 및 내재해성 인삼의 분자육종을 비교적 단시간내에 할 수 있는 형질전환 및 재분화 시스템을 개발하고 토양순화 시스템을 확립하고자 하였다.

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Determination of Complete Genome Sequence of Korean Isolate of Potato virus X

  • Choi, Sun-Hee;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • 제24권3호
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    • pp.361-364
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    • 2008
  • The complete nucleotide sequences of a Korean isolate of Potato virus X(PVX-Kr) has been determined. Full-length cDNA of PVX-Kr has been directly amplified by long template reverse transcription and polymerase chain reaction(RT-PCR) using virus specific 5'-end primer and 3'-end primer, and then constructed in a plasmid vector. Consecutive subclones of a full-length cDNA clone were constructed to identify whole genome sequence of the virus. Total nucleotide sequences of genome of PVX-Kr were 6,435 excluding one adenine at poly A tail, and genome organization was identical with that of typical PVX species. Comparison of whole genome sequence of PVX-Kr with those of European and South American isolates showed 95.4-96.8% and 77.4-77.9%, in nucleotide similarity, respectively. Sequenced PVX-Kr in this study and twelve isolates already reported could be divided into two subgroups in phylogeny based on their complete nucleotide sequences. Phylogenetic tree analysis demonstrated that PVX-Kr was clustered with European and Asian isolates(Taiwan, os, bs, Kr, S, X3, UK3, ROTH1, Tula) in the same subgroup and South American isolates(CP, CP2, CP4, HB) were clustered in the other subgroup.

구두충(Longicollum pagrosomi)에 감염된 참돔(Pagrus major)의 Heat Shock Protein 70 (Hsp70) 발현 (Expression of a Heat Shock Protein 70 (Hsp70) in Red Seabream Pagrus major Infected with Longicollum pagrosomi)

  • 박형준;민병화
    • 한국수산과학회지
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    • 제51권2호
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    • pp.163-169
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    • 2018
  • This study examined the expression of heat shock protein 70 (Hsp70) in red seabream Pagrus major infected by the, acanthocephalan parasites Longicollum pagrosomi. We cloned the full-length Hsp70 cDNA from the liver of the red seabream. The full-length cDNA had a 1,950 bp open reading frame (ORF) that encoded a protein of 650 amino acids. The deduced amino acid sequence of Hsp70 contained all of the conserved Hsp70 family signature sequences and an adenosine triphosphate (ATP)/guanosine triphosphate (GTP) binding motif, including the EEVD (consensus sequence that terminates in Hsp70 family) consensus sequence. The expression of Hsp70 mRNA was upregulated int the fish head-kidney and liver, as determined by quantitative real-time PCR. We quantified the Hsp70 mRNA expression in normal red seabream and fish infected fish by L. pagrosomi. The expression of Hsp70 mRNA was significantly higher in the infected red seabream. These results suggest that Hsp70 play a role of protection against stress and inflammation caused by the parasite and may help maintain homeostasis.

Genomic Organization of ancop Gene for ${\alpha}-COP$ Homolog from Aspergillus nidulans

  • Lee, Hwan-Hee;Chae, Shun-Kee;Kim, Jeong-Yoon;Maeng, Pil-Jae;Park, Hee-Moon
    • Mycobiology
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    • 제28권4호
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    • pp.171-176
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    • 2000
  • We have cloned a ${\alpha}-COP$ homolog, ancop, from Aspergillus nidulans by colony hybridization of chromosome specific library using ${\alpha}-COP$ homologous fragment as a probe. The probe DNA was amplified with degenerated primers designed by comparison of conserved region of the amino acid sequences of Saccharomyces cerevisiae ${\alpha}-COP$, Homo sapiens HEP-COP, and Drosophila melanogaster ${\alpha}-COP$. Full length cDNA clone was also amplified by RT-PCR. Comparison of genomic DNA sequence with cDNA sequence obtained by RT-PCR revealed 7 introns. Amino acid sequence similarity search of the anCop with other ${\alpha}-COPs$ gave an overall identity of 52% with S. cerevisiae, 47% with human and bovine, 45% with Drosophila and Arabidopsis. In upstream region from the transcription start site, a putative TATA and CAAT motif were also identified.

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