• 한국동물번식학회:학술대회논문집
• /
• 한국동물번식학회 2004년도 춘계학술발표대회
• /
• pp.211-211
• /
• 2004

골다공증 치료제로 이용되고 있는 hPTH (human parathyroid hormome)는 체내의 혈중 칼슘 농도를 조절하는 인간의 부갑상선 호르몬이다. 본 연구에서는 retrovirus vector system을 이용하여 hPTH를 효율적으로 생산하는 PFF (porcine fetal fibroblast) 돼지 세포를 구축하고자 하였다. hPTH 유전자는 갑상선 암 환자로부터 적출한 부갑상선 조직의 RNA를 주형으로 RT-PCR을 수행하여 cloning하였으며, 이 유전자가 RSV promoter 통제하에 발현되게끔 design된 retrovirus vector를 구축하였다. (중략)

• 한국미생물·생명공학회지
• /
• 제14권2호
• /
• pp.155-160
• /
• 1986

E. coli-Yeast shuttle vector YEp 13에 B. amyloliquefaciens의 $\alpha$-amylase gene을 cloning하여 얻은 hybrid plasnidlasmid를 E. coli를 숙주세포로 하여 형질을 발현시켰다. 형질전환주의 $\alpha$-amylase 활성 측정 결과, B. amyloliquefaciens에 대해 20-30%의 상대 활성도를 나타내었다. 형질전환주의 경우 생성된 $\alpha$-amylase의 60-65%가 periplasm에 축적되었으며 세포 외부로의 분비는 없었다. Hybrid DNA를 agarose-gel 전기영동으로 조사한 결과 그 크기가 다른 다수의 hybrid DNA가 확인되었으며 pHA28의 plasmid (a)(Fig.3)에서만 $\alpha$-amylase 활성을 나타내는 것으로 밝혀졌다 pHA28의 plasmid(a)의 크기가 YEp 13 plasmid DNA보다 작은 것은 YEp13 plasmid에서 yeast gene부분이 deletion된 결과로 추측되었다.

• 미생물학회지
• /
• 제30권4호
• /
• pp.246-251
• /
• 1992

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.

• Journal of Microbiology
• /
• 제44권6호
• /
• pp.671-673
• /
• 2006

A new 4.87 kb Escherichia-Pseudomonas shuttle vector has been constructed by inserting a 1.27 kb DNA fragment with a replication origin of a Pseudomonas plasmid pRO1614 into the 3.6 kb E. coli plasmid pBGS18. This vector, designated pJH1, contains an aminogly-coside phosphotransferase gene (aph) from Tn903, a lacZ' gene for $\alpha$-complementation and a versatile multiple cloning site possessing unique restriction sites for EcoRI, SacI, KpnI, SmaI, BamHI, XbaI, SalI, BspMI, PstI, SphI, and HindIII. When pJH1 was transformed into E. coli DHS${\alpha}$ and into P. putida HK-6, it was episomally and stably maintained in both strains. In addition, the enhanced green fluorescent protein (EGFP) gene which was transcriptionally cloned into pJH1 rendered E. coli cells fluorescence when its transformants were illuminated at 488 nm.

• Journal of Microbiology and Biotechnology
• /
• 제1권4호
• /
• pp.240-245
• /
• 1991

We have constructed a promoter-probe vector pKU20 using pKT230, a derivative of broad-host-range plsmid RSF1010, as a base. The pKU20 contains structural gene for aminoglycoside phos-photransferase (aph), without promoter, and a multiple cloning site upstream the aph. Using this vector, a 412base pairs (bp) PstI fragment showing strong promoter activity both in Escherichia coli LE392 and Pseudomonas putida KCTC1644 has been cloned from Pseudomonas fluorescens chromosomal DNA on the basis of streptomycin resistance. The nucleotide sequence of the 412 bp fragment has been determined and the putative - 35 and -10 region was observed. Insecticidal protein gene of Bacillus thuringiensis subsp. kurstaki HD-73 inserted on downstream of the promoterlike DNA fragment was efficiently expressed in E. coli and P. putida. The toxin protein was efficiently synthesized in an insoluble form in both strains.

• KSBB Journal
• /
• 제5권1호
• /
• pp.49-58
• /
• 1990

대장균으로부터 alpha interferon의 생산과 분비를 유도하기 위해 대장균의 lipoprotein promoter, lactose promoter 및 operator와 lipoprotein의 signal seqquence를 가지는 vector에 alpha-IFN유전자를 cloning하여 발현 vector pIF-III-B와 vector pIF-III-C를 제작하였다.

• 한국미생물·생명공학회지
• /
• 제29권1호
• /
• pp.1-11
• /
• 2001

Lactic acid bacteria (LAB) are widely used for various food fermentation. With the recent advances in modern biotechnology, a variety of bio-products with the high economic values have been produced using microorganisms. For molecular cloning and expression studies on the gene of interest, E. coli has been widely used mainly because vector systems are fully developed. Most plasmid vectors currently used for E, coli carry antibiotic-resistant markers. As it is generally believed that the antibiotic resistance markers are potentially transferred to other bacteria, application of the plasmid vectors carrying antibiotic resistance genes as selection markers should be avoided, especially for human consump-tion. By contrast, as LAB have some desirable traits such that the they are GRAS(generally recognized as safe), able to secrete gene products out of cell, and their low protease activities, they are regarded as an ideal organism for the genetic manipulation, including cloning and expression of homologous and heterologous genes. However, the vec-tor systems established for LAB are stil insufficient to over-produce gene products, stably, limiting the use of these organisms for industrial applications. For a past decade, the two popular plasmid vectors, pAM$\beta$1 of Streptococcus faecalis and pGK12 theB. subtilis-E. coli shuttle vector derived from pWV01 of Lactococcus lactis ssp. cremoris wg 2, were most widely used to construct efficient chimeric vectors to be stably maintained in many industrial strains of LAB. Currently, non-antibiotic markers such as nisin resistance($Nis^{r}$ ) are explored for selecting recombi-nant clone. In addition, a gene encoding S-layer protein, slp/A, on bacterial cell wall was successfully recombined with the proper LAB vectors LAB vectors for excretion of the heterologous gene product from LAB Many food-grade host vec-tor systems were successfully developed, which allowed stable integration of multiple plasmid copies in the vec-mosome of LAB. More recently, an integration vector system based on the site-specific integration apparatus of temperate lactococcal bacteriophage, containing the integrase gene(int) and phage attachment site(attP), was pub-lished. In conclusion, when various vector system, which are maintain stably and expressed strongly in LAB, are developed, lost of such food products as enzymes, pharmaceuticals, bioactive food ingredients for human consump-tion would be produced at a full scale in LAB.

• 문화기술의 융합
• /
• 제8권3호
• /
• pp.469-475
• /
• 2022

딥러닝 기반 1인 화자 TTS 시스템의 모델을 학습하기 위해서 수십 시간 분량의 음성 DB와 많은 학습 시간이 요구된다. 이것은 다화자 또는 개인화 TTS 모델을 학습시키기 위해서는 시간과 비용 측면에서 비효율적 방법이다. 음색 복제 방법은 새로운 화자의 TTS 모델을 생성하기 위하여 화자 인코더 모델을 이용하는 방식이다. 학습된 화자 인코더 모델을 통해 학습에 사용되지 않은 새로운 화자의 적은 음성 파일로부터 이 화자의 음색을 대표하는 화자 임베딩 벡터를 만든다. 본 논문에서는 음색 복제 방식을 적용한 다화자 TTS 시스템을 제안한다. 제안한 TTS 시스템은 화자 인코더, synthesizer와 보코더로 구성되어 있는데, 화자 인코더는 화자인식 분야에서 사용하는 d-vector 기법을 적용한다. 학습된 화자 인코더에서 도출한 d-vector를 synthesizer에 입력으로 추가하여 새로운 화자의 음색을 표현한다. MOS와 음색 유사도 청취 방법으로 도출한 실험 결과로부터 제안한 TTS 시스템의 성능이 우수함을 알 수 있다.

• BMB Reports
• /
• 제38권2호
• /
• pp.191-197
• /
• 2005

Although the human genome has been nearly completely sequenced, the functions and the roles of the vast majority of the genes, and the influences of single nucleotide polymorphisms (SNPs) in these genes are not entirely known. A modified mutation detection method was developed for large-scale cloning of the possible SNPs between tumor and normal cells for facilitating the identification of genetic factors that associated with cancer formation and progression. The method involves hybridization of restriction enzyme-cut chromosomal DNA, cleavage and modification of the sites of differences by enzymes, and differential cloning of sequence variations with a designed vector. Experimental validations of the presence and location of sequence variations in the isolated clones by PCR and DNA sequencing support the capability of this method in identifying sequence differences between tumor cells and normal cells.

• Mycobiology
• /
• 제37권3호
• /
• pp.240-242
• /
• 2009

An improved procedure for preparing PCR cloning vectors was developed. This procedure includes the incorporation of adapters to create XcmI restriction enzyme sites in pBluescript II SK(+) vectors, digestion with XcmI followed by further digestion of the small fragment produced by XcmI digestion with additional enzymes, and purification with PCR purification kits. Using this procedure, PCR cloning vectors with high ligation efficiencies and low blue or false-positive colonies were obtained.