• Korean Journal of Fisheries and Aquatic Sciences
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• v.46 no.6
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• pp.901-906
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• 2013

We examined the availability of the lacZ gene (${\beta}$-galactosidase gene) as a reporter of foreign gene transfer in the cysts of Artemia franciscana (A. franciscana) to conduct a risk assessment of living genetically modified organisms (LMOs) in the marine ecosystem. The LacZ gene was transferred to decapsulated cysts by particle bombardment, and its insertion and expression were assessed by means of polymerase chain reaction (PCR) and X-gal staining. X-gal staining indicated lacZ expression in all A. franciscana examined (including the control group), which exhibited not only negative but also positive PCR amplification. Endogenous ${\beta}$-galactosidase is highly active in the whole body of A. franciscana during all stages of the life cycle. Thus, the lacZ gene is unsuitable as a reporter for foreign gene transfer in A. franciscana cysts, because it is difficult to discriminate between exogenous and endogenous ${\beta}$-galactosidase activity.

• Proceedings of the Microbiological Society of Korea Conference
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• 2002.10a
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• pp.129-131
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• 2002

• Biomedical Science Letters
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• v.17 no.3
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• pp.185-190
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• 2011

Adenovirus type 5 (Ad5) vectors have been used for gene transfer to a wide variety of cell types in vivo and in vitro. The advantages of adenovirus vectors include the high titer of virus readily obtained in large scale preparations, their ability to transduce dividing and non dividing cells, and the high level of transgene expression. Since adenovirus vectors do not integrate in host cell DNA, there is a lack of insertional mutagenesis. However, many human tumor cells lack expression of the adenovirus 5 receptors and contain areas of hypoxia. In order to identify the pattern of replication and gene expression of oncolytic adenovirus in hypoxic condition, multiple different fiber modified Ads (Ad5F/S11, Ad5F/S35, Ad5F/K7, Ad5F/K21, and Ad5F/RGD) was compared. The replication of all fiber modified adenovirus was inhibited in hypoxic condition in HEK 293 cells, but gene expression has variety on different tumor cell lines and the level of coxackievirus and adenovirus receptor (CAR) expression. These data suggest that CAR expression pattern and hypoxic condition of tumor are considered for optimal oncolytic adenovirus application.

• Journal of Microbiology and Biotechnology
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• v.15 no.6
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• pp.1346-1352
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• 2005

We have developed a modified gene replacement method using PCR products containing short homologous sequences of 40- to 50-nt. The method required $\lambda$-Red recombination functions provided under the control of a temperature-sensitive CI857 repressor expressed from the $P_{lac}$ promoter in the presence of IPTG on an easily curable helper plasmid. The method promoted the targeted gene replacements in the Escherichia coli chromosome after shifting cultures of the recombinogenic host, which carries the helper plasmid, to $42^{\circ}C$ for 15 min. Since this method employs $\lambda$-Red recombination functions expressed from the easily curable helper plasmid, multiple rounds of gene replacements in the E. coli chromosome would be possible. The procedures described herein are expected to be widely used for metabolic engineering of E. coli and other bacteria.

• BMB Reports
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• v.41 no.10
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• pp.739-744
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• 2008

Turnip yellow mosaic virus (TYMV) is a positive strand RNA virus that infects mainly Cruciferae plants. In this study, the TYMV genome was modified by inserting an extra subgenomic RNA promoter and a multiple cloning site. This modified TYMV was introduced into Nicotiana benthamiana using a Agrobacterium-mediated T-DNA transfer system (agroinfiltration). When a gene encoding $\beta$-glucuronidase or green fluorescent protein was expressed using this modified TYMV as a vector, replication of the recombinant viruses, especially the virus containing $\beta$-glucuronidase gene, was severely inhibited. The suppression of replication was reduced by co-expression of viral silencing suppressor genes, such as tombusviral p19, closteroviral p21 or potyviral HC-Pro. As expected, two subgenomic RNAs were produced from the recombinant TYMV, where the larger one contained the foreign gene. An RNase protection assay revealed that the recombinant subgenomic RNA was encapsidated as efficiently as the genuine subgenomic RNA.

• KSBB Journal
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• v.11 no.4
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• pp.431-437
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• 1996

The nar promoter of Escherichta coli, which is maximally induced under anaerobic conditions in the presence of nitrate, was characterized to see whether the nar promoter cloned onto pBR322 can be used as an expression promoter. The modified nar promoter, in which several bases in the -10 region was mutated to the consensus sequence by site-directed mutagenesis, was characterized in E. coli, on which chromosome the fnr gene affecting expression of the nar promoter according to dissolved oxygen level was mutated. The E. coli lacZ gene was used as a reporter gene. The following effects were investigated to find optimal conditions to induce the modified nar promoter: induction methods, optimal nitrate concentrations, the amount of ${\beta}$-galactosidase expressed at the different growth conditions, and induction characteristics. The following results were obtained from the experiments : expression of ${\beta}$-galactosidase from the modified nar promoter was not affected much by nitrate concentrations. The maximal specific ${\beta}$-galactosidase activity was obtained when E. coli was grown under aerobic conditions, and then the modified nar promoter was induced at OD600=2.2 under microaerobic conditions (DO=1∼2%), under which conditions the maximal specific ${\beta}$-galactosidase activity was 13,000 Miller units. However, the specific ${\beta}$-galactosidase activity was approximately 6,000 Miller units even before the modified nar promoter was induced. Therefore, the modified nar promoter seemed to be useful when the cloned gene wants to be expressed in E. coli constitutively.

• Animal cells and systems
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• v.15 no.3
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• pp.243-249
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• 2011

The genetic diversity and population history of the Asian shore crab, Hemigrapsus sanguineus, were investigated with a nucleotide sequence analysis of 536 base pairs (bp) of the mitochondrial cytochrome c oxidase subunit I gene (COI) in 111 samples collected from four populations in Korea and one in Japan. In total, 28 haplotypes were defined by 27 variable nucleotide sites in the COI region examined. The observed haplotypes had a shallow haplotype genealogy and no geographical associations. Most of the populations had high haplotype diversity (0.656-0.788) and low nucleotide diversity (0.00165-0.00244), and significant negative values for Fu's $F_S$, suggesting rapid and recent population growth from an ancestral population and sudden population expansion. The pairwise fixation indices ($F_{ST}$) estimated with the exact test and the migration rates indicate that substantial gene flow occurs among these populations as a result of sea currents, except between the Yellow Sea coast of Korea (BUA) and the Pacific Ocean coast of Japan (JPA). These two populations (BUA and JPA) showed significant genetic differentiation and low migration rate.

• Fisheries and Aquatic Sciences
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• v.14 no.1
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• pp.16-21
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• 2011

The genetic diversity and population genetic structure of thread-sail filefish, Stephanolepis cirrhifer (Temminck & Schlegel), were examined with a nucleotide sequence analysis of a 495bp fragment of the 5'-end of the cytochrome b gene in 113 fish collected from five populations from the south and east coasts of the Korean Peninsula. Seventeen variable nucleotide sites and 16 haplotypes were defined. The observed haplotypes had a shallow haplotype genealogy and no geographical association. Most of the populations had high haplotype diversity and low nucleotide diversity, and significant negative values for Fu's $F_S$, suggesting rapid, recent population growth from an ancestral population and sudden population expansion. The estimated pairwise fixation indices ($F_{ST}$) indicate that substantial gene flow occurs among these populations. Thread-sail filefish in the South Sea of Korea and East Sea Korean populations forms a single panmictic population. Thus, thread-sail filefish in these areas should be treated as one management unit.

• The Korean Journal of Applied Statistics
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• v.2 no.1
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• pp.35-47
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• 1989

This article deals with the method of ML among the methods of estimating m gene frequenecies in the Generalized ABO-like Blood Group Systems and with the statistical testing about the differencies of gene frequencies by using these estimators. Especially, the generalization about the Homogeneity testing problem is tried and thus it enables us to test of Homogeneity of m gene frequencies. Finally, in the example, ML estimator is compared with other estimators suggested by Bernstein method, by adjusted Bernstein method and by modified Bernstein method, and statistical testing in the above is carried out by using orthogonal partitioning.

• Fisheries and Aquatic Sciences
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• v.16 no.4
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• pp.243-251
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• 2013

Genetic diversity and gene flow patterns in Pollicipes mitella were investigated with a nucleotide sequence analysis of 514 base pairs from the mitochondrial cytochrome c oxidase subunit I gene (COI) in 124 samples collected from six Korean populations. In total, 59 haplotypes were defined by 40 variable nucleotide sites in the COI region. The haplotypes had shallow haplotype genealogy and no geographic associations. All populations had high haplotype diversity (0.909 to 0.979) and low nucleotide diversity (0.0055 to 0.0098). The haplotypes with recently diverged nucleotides were distributed by long-range larvae dispersal among regional populations. The pairwise fixation indices ($F_{ST}$) estimated with the exact test and migration rates indicate that substantial gene flow has occurred among populations as a result of sea currents, except between the Uljin (East Sea coast) and other Korean populations. This suggests that significant genetic differentiation and low migration rates have affected the Uljin population.