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Cloning 된 효모의 RNAI 유전자의 특성에 관하여
김진경,김대영,송영환 한국어병학회 1993 한국어병학회지 Vol.6 No.2
효모의 RNAI유전자는 RNA processing에 관여 하는지 혹은 RNA transport에 관여 하는지 아직까지 유전자의 기능이 정확히 알려져 있지 않은 실정이다. 효모의 RNAI 유전자의 기능을 파악하기 위한 방법으로 본 연구에서는 rnal- 1 mutant gene을 cloning하여 이에 대한 DNA sequence를 조사함으로써 RNAI 유전자와 rnal-1 유전자의 차이점을 이해하고자 하였다. mal-1 marker를 갖는 yeast strain(R49)로 부터 genomic DNA를 추출하여 이를 BglII로 절단하여 genomic southern blotting을 행한 결과 wild type의 경우와 동일하게 3.4 kb에서 hybridization되는 signal을 얻었으며, RNAl 및 rnal-1이 yeast genome내에 single site로 존재함을 보여 주는 결과를 얻었다. mutant strain으로 부터 얻은 3.4 kb의 BglI fragment를 pUC19의 BamHI site에 subcloning하여 transformant들을 얻었고, wild type RNAl 유전자를 probe로 하여 rnal-1 mutant 유전자를 cloning할 수 있었다. pUC19에 cloning된 RNAl 유전자 및 rnal-1유전자로부터 다양한 Ba131유도체를 얻어 이들에 대한 염기 서열을 비교한 결과 transcription initation site에서부터 down stream쪽으로 17 아미노산위치에 TCC가 TTC로 대치되어 있었으며 그 결과 serine이 phenylalanine으로 변환되는 결과를 얻었다. Wild type RNAI gene의 5'-region에는 3군데의 TATA-like sequence가 true TATA box인지 확인하기 위하여 Bal3I deletion에 의해 -103nt까지 deletion된 유도체를 얻었으며 △RNAI, rnal-1, 81-2-6 clone이 rnal-1 allele와 complementation한지 확인하였으나 △ RNAI은 TS-complementation을 하지 못하였다. 따라서 현재까지 TATA-box라고 알려진 부분은 promoter로 작용하지 못함을 확인하였다. The RNAI mutation of Saccharomyces cerevisia is a recessive and temperature sensitive lethal mutation which interferes with the production of mRNA, rRNA, and tRNA. However, the precise role of RNAI gene have not been revealed until yet. We have cloned rnal-1 mutant gene from rnal-1 mutant yeast strain(R49 ; trpl, ura3-52, rnal-1). The 3.4kb BglII fragment of wild type RNAI clone(81-2-6) contains whole RNAI gene. The genomic southern blotting with BglII digested R49 genomic DNA as a probe shows the unique and identical band with wild type 3.4kb BglII fragment. Therefore, We prepared partial BglII genomic library(3∼4kb BglII fragments) into BamH I site of pUC19. The rna 1-1 mutant clone was screened with Digoxigenin(DIG)-lableled probe by high density colony hybridization. The 5'-flanking region of rnal-1 gene was sequenced by dideoxy chain termination method. The 5'-flanking sequence of RNAI gene contains three TATA-like sequence ; TAATA, TATA and TTTTAA at position of -67, -45, and -36 from first ATG codon respectively. The 5'-flanking region of wild type RNAI gene from ATG codon to -103nt was deleted with Bal31 exonuclease digestion, generating pUC△/RNAI. After constructing pYEP△RNA I (consists of -103nt deleting RNAI gene, URA3 gene, 2㎛ rep. origin), pYEPrnal-1(consists of Xba I fragment of pUCrnal-1. URA3 gene, 2㎛ rep. origin), and pYEPRNAI. each plasmid was transformed into host strain(trpl, ura3-52, rnal-1) by electroporation, respectively. Yeast transformant carrying pYEP△RNAI did not complement the thermal sensitivity of rnal-1 gene. It means that TATA-like sequences in 5'-flanking region is not TATA sequence for transcribing RNAI gene and there may be other essential sequence in upstream region for the transcription of RNAI gene.
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흡연행태에 따른 Porphyromonas gingivalis의 발현율과 유전형 차이
김진경,Kim, Jin-Kyoung 한국임상보건과학회 2020 한국임상보건과학회지 Vol.8 No.2
Purpose: The purpose of this study was to differences in the expression rate of Porphyromonas gingivalis according to smoking status, smoking amount and period of smoking. Methods: At the time of investigation, 30 smokers and non-smokers were recruited among patients with periodontitis with a probing pocket depth(PPD) of 4 mm or more. General information was collected using a self-questionnaire, and the average value was used by a dentist to measure the probing pocket depth of three times each for the first or second molar. Plaque collection and analysis were performed by collecting only subgingival plaque using a conventional method, and the expression rate of P. gingivalis was confirmed using polymerase chain reaction (PCR). For statistical analysis, the SPSS Ver 25.0 program was used. Results: Smoking did not have a significant effect on the expression of P. gingivalis, but it did affect the expression of more type II genotypes (p<0.05). In addition, smokers had more slight periodontal pocket, and the amount and duration of smoking did not affect the expression of P. gingivalis. Conclusions: In the future, it is necessary to reinforce the group of smokers and non-smokers with healthy oral conditions, and to investigate the quantitative difference in the expression rate and genotype of P. gingivalis over time of harmful substances in smoking.
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김진경,심창기,박상원,박병준,지형진,김원일,권오경,임건재,Kim, Jin-Kyoung,Shim, Chang-Ki,Park, Sang-Won,Park, Byung-Jun,Jee, Hyeong-Jin,Kim, Won-Il,Kwon, Oh-Kyung,Im, Geon-Jae 한국유기농업학회 2009 韓國有機農業學會誌 Vol.17 No.4
시판 마요네즈를 이용하여 오이 흰가루병에 대한 방제효과를 시험하였다. 마요네즈를 0.5~2% 농도로 물에 희석하여 흰가루병이 발병된 오이에 6일 간격 3회 처리했을 때 8.3~99.2% 방제효과를 나타내었다. 90% 이상의 방제효과를 보인 마요네즈 희석액 중 2-3엽의 어린모에서 약해가 없었던 농도는 0.5%이었다. 마요네즈 중 기름함량이 78~80% 수준인 일반 마요네즈와 기름함량을 38~70%로 줄인 저지방 마요네즈의 흰가루병 방제력을 비교했을때 저지방 마요네즈는 방제율은 39.3~59.8% 수준으로 일반마요네즈에 비해 방제율이 떨어졌다. 마요네즈가 처리된 잎의 엽록소 함량은 발병구에 비해 엽록소 함량이 약 3배 정도 높게 조사되어 처리된 마요네즈가 오이 잎의 광합성을 저해하지 않으면서 흰가루병을 효과적으로 방제함을 확인하였다. This study was conducted to develop an organic control of powdery mildews of cucumber by using mayonnaise in green house. The treatment of 0.1~2% mayonnaise resulted in 8.3%~99.2% control efficiencies against powdery mildew of cucumber. 0.5% mayonnaise treatment resulted control values over 97% in disease. It did not adversely affected the photosynthesis of foliages. Although one application of mayonnaise to the foliage was not practically effective enough, two or three application of mayonnaise to the foliage at the 0.5% concentration resulted in excellent control against powdery mildews. This treatment could provide protection for 10~14 days after application. Among the type of mayonnaise, general mayonnaise revealed 97.5% control value, but mayonnaise containing low oil content revealed 39.3%~97.5% control values on powdery mildews at the 0.5% concentration. Therefore, oil content in mayonnaise played a essential material to control powdery mildew. Results indicated that mayonnaise could be used as organic control of powdery mildews of cucumber. This control might be environmentfriendly as well as cost-effective.
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