• Journal of Microbiology and Biotechnology
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• v.26 no.12
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• pp.2184-2191
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• 2016

The time-consuming and high-cost preparation of soluble peptide-major histocompatibility complexes (pMHC) currently limits their wide uses in monitoring antigen-specific T cells. The single-chain trimer (SCT) of peptide-${\beta}2m$-MHC class I heavy chain was developed as an alternative strategy, but its gene fusion is hindered in many cases owing to the incompatibility between the multiple restriction enzymes and the restriction endonuclease sites of plasmid vectors. In this study, overlap extension PCR and one-step cloning were adopted to overcome this restriction. The SCT gene of the $OVA_{257-264}$ peptide-$(GS_4)_3-{\beta}2m-(GS_4)_4-H-2K^b$ heavy chain was constructed and inserted into plasmid pET28a by overlap extension PCR and one-step cloning, without the requirement of restriction enzymes. The SCT protein was expressed in Escherichia coli, and then purified and refolded. The resulting $H-2K^b/OVA_{257-264}$ complex showed the correct structural conformation and capability to bind with $OVA_{257-264}$-specific T-cell receptor. The overlap extension PCR and one-step cloning ensure the construction of single-chain MHC class I molecules associated with random epitopes, and will facilitate the preparation of soluble pMHC multimers.

• KSII Transactions on Internet and Information Systems (TIIS)
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• v.2 no.2
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• pp.120-133
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• 2008

This paper presents a novel approach for facial motion tracking and facial expression cloning to create a realistic facial animation of a 3D avatar. The exact head pose estimation and facial expression tracking are critical issues that must be solved when developing vision-based computer animation. In this paper, we deal with these two problems. The proposed approach consists of two phases: dynamic head pose estimation and facial expression cloning. The dynamic head pose estimation can robustly estimate a 3D head pose from input video images. Given an initial reference template of a face image and the corresponding 3D head pose, the full head motion is recovered by projecting a cylindrical head model onto the face image. It is possible to recover the head pose regardless of light variations and self-occlusion by updating the template dynamically. In the phase of synthesizing the facial expression, the variations of the major facial feature points of the face images are tracked by using optical flow and the variations are retargeted to the 3D face model. At the same time, we exploit the RBF (Radial Basis Function) to deform the local area of the face model around the major feature points. Consequently, facial expression synthesis is done by directly tracking the variations of the major feature points and indirectly estimating the variations of the regional feature points. From the experiments, we can prove that the proposed vision-based facial expression cloning method automatically estimates the 3D head pose and produces realistic 3D facial expressions in real time.

• Journal of Microbiology and Biotechnology
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• v.6 no.2
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• pp.142-144
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• 1996

To clone the gene coding for steroid $\Delta^1$-dehydrogenase of Arthrobacter simplex, its genomic library was constructed with a , $\lambda$gt11 expression vector and immunoscreened with antiserum against the enzyme. One positive clone was found to carry a 1.6-kb EcoR I restriction endonuclease fragment of A. simplex DNA. The restriction map of the 1.6-kb EcoR I fragment was determined after cloning of the DNA into pBS vector.

• The Journal of Information Technology
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• v.12 no.2
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• pp.1-12
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• 2009

China first started its mobile phone services in 1987, and the number of users has exponentially increased so that it reached 700 millions in January 2009. Currently China's 2G(GSM) users is 650 millions. These 2G (GSM) services have an advantage of the capability to use the mobile phone with a SIM (Subscriber Identity Modul) card, one kind of smart cards, inserted into it. However, due to the security vulnerability of SIM cards being used within China's 2G (GSM) services, SIM cards cloning. Problems concerning mobile phone surveillance towards a designated person by illegal cloning ESN and IMSI have recently risen to be a massive social issue within Korea as well. These studies have experimented the possibility of SIM cards clone in various mobile communication corporations using 2G (GSM) in China, and hence discovered the security vulnerability such as the incoming outgoing, SMS service and additional services on mobile phones using clone SIM cards.

• Korean Journal of Environmental Biology
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• v.21 no.1
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• pp.56-59
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• 2003

The Salmonella typhimurium cdd gene encoding cytidine deaminase (cyti-dine/2'-deoxycytidine aminohydrolase; EC 3.5.4.5.) was isolated through shotgun clon-ing by complementation of the E. coli odd mutation. By subsequent deletion and sub-cloning from the original 3.7 Kb of EcoRI insert (pSAMI), the precise region of the cdd structural gene is located around the BglII site in the middle part of 1.7 Kb of NruI/PvuI segment. The 1.7 Kb containing odd gene wag subcloned to the pUC18 vector and the nucleotide sequence of the cdd gene was determined. When the putative ribosorne-binding site (Shine-Dalgarno sequence) and initiation codon were predicted to be GAGG at the position 459 and ATG at the position 470, respectively, there was an open reading frame of 885 nucleotides, encoding an 294 amino acid protein. The cdd gene expression in E. coli JF611/pSAMI was amplified about 50 fold compared to that of the wild type. The cdd gene expression was maintained in the stationary phase after rea-ching the peak in the late logarithmic phase.

• Applied Biological Chemistry
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• v.31 no.4
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• pp.371-375
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• 1988

Streptomycetes are attractive microorganisms for their production of various secondary metabolites such as antibiotics. Now, the development of gene manipulation in this microorganisms enables the cloning and analysis of the genes which coding for antibiotic biosynthesis and resistance to the drug. In this article, we reviewed the studies with respect to the mechanisms of self-protection and cloning of the genes cloning for antibiotic biosynthesis, particularly, in microorganisms which produce antibiotic inhibitors of protein synthesis.

• Journal of Microbiology and Biotechnology
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• v.1 no.4
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• pp.251-255
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• 1991

A 16 kilobase (kb) HindIII fragment of alkalophilic Bacillus sp. YC-335 containing a gene for xylanase synthesis was inserted at the HindIII site of pBR322 and cloned in Escherichia coli HB101. After subcloning of recombinant plasmid pYS52, the 1.5 kb fragment was found to code for xylanase activity, and the hybrid plasmid was named pYS55. The DNA insert of the plasmid was subjected to restriction enzyme mapping, which showed that pYS55 had single site for PuvII and SstI in the 1.5 kb insert fragment. Southern hybridization analysis revealed that the cloned gene was hybridized with chromosomal DNA from alkalophilic Bacillus sp. YC-335. About 64% of the enzyme activity was observed in the extracellular and periplasmic space of E. coli HB10l carrying pYS55.

• Archives of Pharmacal Research
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• v.12 no.3
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• pp.176-180
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• 1989

Four bacterial strains having inducible resistance to erythromycin were isolated from soil samples in Korea and characterized. MLS inducibility was checked in each strain. Cloning of inducible resistance gene(s) has been tried. Four isolates were identified as B. anthracis, Micrococcus luteus, Streptococcus faecium and B. licheniformis, in which erythromycin, oleandomycin, cirramycin and carbomycin acted as resistance inducers respectively. The resistance gene cloned from B, licheniformis 597 strain using pBS 42 vector was found to have a 3.2 kb insert.

• Journal of the Korean Society of Tobacco Science
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• v.23 no.2
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• pp.103-108
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• 2001

Hog cholera virus(HCV) was purified from virus infected Bovine kidney cells. From this virus, total protein was analyzed by SDS-PAGE gel electrophoresis and about 55 kDa band of E2 envelope protein was detected. The viral RNA was purified and E2 cDNA was amplified by RT-PCR. E2 cDNA fragment was cloned to PCRII-TOPO cloning vector and named pE2. The analysis of nucleotide sequence showed that this E2 cDNA fragment inserted into pE2 was 1191 nucleotides long and coded 397 amino acids.

• Korean Journal of Microbiology
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• v.35 no.1
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• pp.25-27
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• 1999

Degenerate primers corresponding to the sequences of the copper-binding regions in the fungal laccases were used to isolatc laccase gene specific fragment by PCR in Coprinus congregahts. A 144 bp DNA hagrnent was cloned and was identified to have 60-69 % homology with other fungal laccase genes. The predicted amino acid sequcnces showed 68-75% homology with other fungal laccase proteins.