The faulty regulation of imprinting gene lead to the abnormal development of reconstructed embryo after nuclear transfer. However, the correlation between the imprinting status of donor cell and preimplantation stage of embryo development is not yet clear. In this study, to determine this correlation, we used the porcine spermatogonial stem cell (pSSC) and fetal fibroblast (pFF) as donor cells. As the results, the isolated cells with laminin matrix selection strongly expressed the GFRα-1 and PLZF genes of SSCs specific markers. The pSSCs were maintained to 12 passages and positive for the pluripotent marker including OCT4, SSEA1 and NANOG. The methylation analysis of H19 DMR of pSSCs revealed that the zinc finger protein binding sites CTCF3 of H19 DMRs displayed an androgenic imprinting pattern (92.7%). Also, to investigate the reprogramming potential of pSSCs as donor cell, we compared the development rate and methylation status of H19 gene between the reconstructed embryos from pFF and pSSC. This result showed no significant differences of the development rate between the pFFs (11.2±0.8%) and SSCs (13.3±1.1%). However, interestingly, while the CTCF3 methylation status of pFF-NT blastocyst was decreased (36.3%), and the CTCF3 methylation status of pSSC-NT blastocyst was maintained. Therefore, this result suggested that the genomic imprinting status of pSSCs is more effective than that of normal somatic cells for the normal development because the maintenance of imprinting pattern is very important in early embryo stage.

The first commercial bioinsecticide in Korea was registered in 2003 to control diamond back moth and beet armyworm in Chinese cabbage using Bacillus thuringiensis subsp. aizawai. Currently, fourteen microbial pesticides have registered in Korean market. The Korean government is aiming for 40% reduction (from the 2004 value) in the use of chemical pesticides by 2013. To increase the use of microbial pesticides as an alternative of chemicals, we conducted in vitro bioassay with Bt products. Bt products were treated alone and in combination against larvae of beet armyworm, diamond back moth and tobacco cutworm. Five Bt products shown high mortality against diamond back moth in recommended concentration, but had low control effects against beet armyworm and tobacco cutworm. Mixed Bt products had no synergic effects against the tested three moths. When Bt products were sprayed with higher concentration than recommended dose, those mortality was similar with lower concentration.

감국(Dendranthema indicum)의 환경조건에 따른 개화기의 초장, 개화소요일수, 카로티노이드 함량 및 peroxidase 활성을 고랭지(대관령, 표고 800m)와 평난 지(강릉, 표고 20m)의 노지와 온실에서 각각 비교하여 원예화 개발 가능성을 검토하였다. 환경조건에 따른 광 량은 고랭지 노지 > 평난지 노지 > 고랭지 온실 > 평난지 온실 순으로 감소하였다. 고랭지의 노지에서는 UV A와 UV B의 세기가 22.4, 3.02W·m-2로 평난지 노지의 19.9, 2.24W·m-2보다 각각 12.6, 34.8% 더 강했다. 감국의 초장은 평난지의 온실조건에서 가장 컸 으나 개화는 고랭지의 노지에서 가장 빨랐다. 감국 꽃 의 주요 색소인 카로티노이드는 고랭지의 온실에서 60.9OD·g-1으로 가장 많았다. 그러나 peroxidase 활성 은 평난지의 온실에서 자란 식물체에서 5.72unit·mg-1 protein 가장 높았다.

Insulin-like growth factor II (IGF2) and H19 genes are mutually imprinted genes which may be responsible for abnormalities in the cloned fetuses and offspring. This study was performed to identify putative differentially methylated regions (DMRs) of porcine H19 locus and to explore its genomic imprinting in in vitro fertilized (IVF) and somatic cell nuclear transferred (SCNT) embryos. Based on mice genomic data, we identified DMRs on H19 and found porcine H19 DMRs that included three CTCF binding sites. Methylation patterns in IVF and SCNT embryos at the 2-, 4-, 8～16-cells and blastocyst stages were analyzed by BS (Bisulfite Sequencing)-PCR. The CpGs in CTCF1 was significantly unmethylated in the 2-cell stage IVF embryos. However, the 4- (29.1%) and 8～16-cell (68.2%) and blastocyst (48.2%) stages showed higher methylation levels (p<0.01). On the other hand, SCNT embryos were unmethylayted (0～2%) at all stages of development. The CpGs in CTCF2 showed almost unmethylation levels at the 2-, 4- and 8～16-cell and blastocyst stages of development in both IVF (0～14.1%) and SCNT (0～6.4%) embryos. At all stages of development, CTCF3 was unmethylated in IVF (0～17.3%) and SCNT (0～1.2%) embryos except at the blastocyst stage (54.5%) of IVF embryos. In conclusion, porcine SCNT embryos showed an aberrant methylation pattern comprised to IVF embryos. Therefore, we suggest that the aberrant methylation pattern of H19 loci may be a reason for increased abnormal fetus after embryo transfer of porcine SCNT embryos.