The canine major histocompatibility complex (MHC) is referred to dog leukocyte antigens (DLA), which is known to be the most polymorphic genetic system in canine species. Many cloned dogs have been produced since Snuppy, first cloned dog, there was no research about genetic identity of MHC among cloned animals. Recently in Lee’s group, two non-transgenic cloned beagles (BG1, 2) were produced by somatic cell nuclear transfer (SCNT) using fetal fibroblast (BF). Also, four transgenic cloned beagles (Ruppy 1-3, 5) were generated using transgenic BF transfected with Red fluorescent protein (RFP) gene. We hypothesize that non-transgenic (BG1, 2) and transgenic (Ruppy 1-3, 5) cloned beagles derived from identical donor cells have the same immunological genetic characteristic except for RFP gene insertion in the genome. Thus, the aim of this study is to confirm the immunological identity of DLA class II in cloned beagles produced using same nuclear donor cell. Genomic DNA was extracted from blood of BG1, BG2, Ruppy 1, 2, 3 and 5. Genomic DNA of normal two control beagle, no correlation with BF was also investigated for rulling out the possibility that beagles were inbred. Forward and reverse primers used for DLA-DQA1 and DQB1 respectively were DQAF: 5’-TAAGGTTCTTTTCTCCCTCT-3’ and DQAR: 5’-GGACAGATTCAGTGAAGAGA-3’ DQBR:5’-CTCACTGGCCCGGCTGTCTC-3’ and DQBR: 5’-CACCTCGC CGCTGCAACGTG-3’. Polymerase Chain Reaction (PCR) products were purified, sequenced directly using the Big Dye Terminator kit. Sequencing analysis was performed on an automated 3730xl DNA analyzer. In experiment 1, sequence of DLA-DQ alpha 1 (DQA1) and DLA-DQ beta 1 (DQB1) exon 2, hypervariabel region, was compared in BG1 and BG2. Experiment 2 also compared the sequence of DQA1 and DQB1 among Ruppy 1, 2, 3 and 5. Experimental 3 compared sequence of DQA1 and DQB1 among all cloned dogs (BG1, BG2 and Ruppy 1-3, 5). As a result, BG1 and BG2 have same allele for DQA1 and DQB1 as we expected. They share DQA1*00101 and DQB1*02901 in experiment 1. In experiment 2, Ruppy 1, 2, 3 and 5 also have identical DQA1*00101 and DQB1*02901 allele. No discrimination between transgenic dogs and cloned dogs was seen in DQA1 and DQB1 Allele in experiment 3. DQA1, DQB1 allele was identified as *00101 and *02901 in all dogs. We provided the allele identity of DQA1and DQB1 in cloned beagles, which can be used as preliminary data for immunological related studies. In conclusion, transgenic cloned dogs despite of red fluorescent protein genes being inserted in their nuclear DNA were immunologically compatible with non-transgenic cloned dogs. We demonstrated that cloned beagles produced using identical nuclear donor were immunologically compatible.

개가시나무 가지의 에탄올 추출물에서 항산화활성을 관찰하였으며, 활성성분을 규명하기 위한 연구를 진행하였다. 그 결과, 4종류의 화합물을 분리하여 동정하였다. 분리 동정된 성분은 catechin(1), epi-catechin(2), tyrosol(3) 및 tiliroside(4)이다. 분리 성분의 항산화활성은 DPPH 라디칼 및 superoxide 음이온 라디칼 소거활성을 이용하여 측정하였다. 화합물 1, 2, 3, 4는 100 µL/mL 농도에서 각각 94.2 %, 93.4 %, 33.6 %, 11.2 %의 DPPH 라디칼 저해활성을 나타내었다. 또한, 화합물 1, 2, 3, 4는 200 µL/mL 농도에서 각각 60.2 %, 35.1 %, 20.6%, 4.5 %의 superoxide 음이온 라디칼 저해활성을 나타내었다. 화합물 1 ∼ 4는 개가시나무에서는 처음으로 분리된 물질이다.

Barley Yellow Mosaic Virus (BaYMV) caused significant reduction in barley yield and is difficult to control due to alive parasitic soil-borne fungus, Palmyra gamines that transmits the, virus. Previous studies have indicated that a virus-free soil could be infested by using virus-contaminated farming machineneries and implements. For the further confirmation of this finding, different proportions of BaYMV-infested soil were mixed into virus free soil. Three barley varieties (Hordum vulgarae, cv "Olbori", "Baegdong" and "Sacheon 6") were sown in pots treated with different rate of P. graminis-infested soil ranging from 0% to 100% in October 20, 2001. Results showed that BaYMV infection increased as the rate of infested soil increased. Initial symptoms were observed in a pots treated with 10% infested soil in all the 3 varieties of barley. "Olbori" had about 5% infection in 20% infested soil and about 10% infection in 40% or 50% infested soil and about 20% infection in 60% infested soil. In "Baegdong", the trend of BYMV occurrence was similar with "Olbori" but the time of severe infection was earlier than "Olbori". BaYMV infection in "Sacheon 6" was even earlier than "Baegdong" with much more severe symptoms than "Baegdong". The growth rate of barley was affected by about 19-22% when grown in 20% infested soil. As the rate of BaYMV infested soil increased the heading date was delayed but the maturing date was early in "Olbori" and "Sacheon 6". Also, reduction rate of culm length in 3 varieties increased with increase of infested soil content. However, "Olbori" showed the highest reduction. "Sacheon 6", have been characterized with long spike length, however was significantly reduced as the infested soil increased. On the other hand, spike length of "Olbori" was not significantly affected despite of increased of infested soil. The reduction rate of 1000 kernel weight was higher in large kernel size cultivar "Sacheon 6" and "Olbori" than small kernel size "Baegdong" as increase of BaYMV-infested soil content.

The aim of this study was to isolate chicken feather-degrading bacteria with high keratinolytic activity and to investigate cultural conditions affecting keratinolytic enzyme production by a selected isolate. A chicken feather-degrading bacterial strain CH3 was isolated from poultry wastes. Isolate CH3 degraded whole chicken feather completely within 3 days. On the basis of phenotypical and 16S rDNA studies, isolate CH3 was identified as Bacillus thuringiensis CH3. This strain is the first B. thuringiensis described as a feather degrader. The bacterium grew with an optimum at pH 8.0 and 37℃, where maximum keratinolytic activity was also observed. The composition of optimal medium for keratinolytic enzyme production was feather 0.1%, sucrose 0.7%, casein 0.3%, K2HPO4 0.03%, KH2PO4 0.04%, MgCl2 0.01% and NaCl 0.05%, respectively. The keratinolytic enzyme had a pH and temperature optima 9.0 and 45℃, respectively. The keratinolytic activity was inhibited ethylenediaminetetraacetic acid, phenylmethylsulfonyl fluoride, and metal ions like Hg2+, Cu2+ and Zn2+. The enzyme activated by Fe2+, dithiothreitol and 2-mercaptoethanol.