The establishment of porcine embryonic stem cells (ESCs) from porcine somatic cell nuclear transfer (SCNT) blastocysts is influenced by in vitro culture day of porcine reconstructed embryo and feeder cell type. Therefore, the objective of the present study was to determine the optimal in vitro culture period for reconstructed porcine SCNT embryos and mouse embryonic fibroblast (MEF) feeder cell type for enhancing colony formation efficiency from the inner cell mass (ICM) of porcine SCNT blastocysts and their outgrowth. As the results, porcine SCNT blastocysts produced through in vitro culture of the reconstructed embryos for 8 days showed significantly increased efficiency in the formation of colonies, compared to those for 7 days. Moreover, MEF feeder cells derived from outbred ICR mice showed numerically the highest efficiency of colony formation in blastocysts produced through in vitro culture of porcine SCNT embryos for 8 days and porcine ESCs with typical ESC morphology were maintained more successfully over Passage 2 on outbred ICR mice-derived MEF feeder cells than on MEF feeder cells derived from inbred C57BL/6 and hybrid B6CBAF1 mice. Overall, the harmonization of porcine SCNT blastocysts produced through in vitro culture of the reconstructed embryos for 8 days and MEF feeder cells derived from outbred ICR mice will greatly contribute to the successful establishment of ESCs derived from porcine SCNT blastocysts.

In our previous studies, the cardiac xenotransplantation from an alpha-1,3-galactosyltransferase knockout pig (GT-MCP-MCP) to cynomolgus monkeys showed a mean survival of 38 days. The objective of this study is to genetically upgrade the GT-MCP-MCP pig, to further enhance membrane cofactor protein (MCP) expression and to express an endothelial specific thrombomodulin (TBM). MCP is a complement regulatory protein and TBM is a coagulation inhibitor. As the dicistronic cassette for wild-type-based MCP and TBM concurrent expressions does not show any increase of MCP, we optimized the MCP codon usage (mMCP) and substituted mMCP for MCP. When the mMCP-TBM cassette was transfected to HeLa cells, we were able to find an increased expression of MCP and endothelial cell-specific TBM expression. The cassette was then transfected into ear-skin fibroblasts isolated from one-month-old #23-4 of a GT-MCP-MCP pig, and the cell populations expressing MCP were obtained by MACS cell sorting. We performed a single cell culture of the selected cells, and obtained clones over expressing 90% MCP. The cells of a clone were used as a donor for nuclear transfer and generated GT-MCP/-MCP/mMCP/TBM pig. The transgenic pig was confirmed to be carrying the cells expressing MCP and functioning as an inhibitor against the cytotoxic effect of normal monkey serum, comparable with donor cells. Thus, we believe that the GT-MCP/-MCP/mMCP/TBM transgenic pig would be potential for the prolongation of xenograft survival in the recipients.

The genetic algorithm (GA), one of the artificial intelligence (AI), is developed based on Darwin's theory of evolution, i.e., the mating of randomly selected objects. If more optimal solution is generated, then it is better to repeat the process of setting the optimum value. In this paper, the method of background music using the genetic algorithm is exploited when the computer game is executed each time. As a result, it has created several music that can be used in the actual game, and it could be confirmed that the other music that is created is different music when performed each time.

Spermatogonial stem cells (SSCs) developed into sperms through spermatogenesis have been utilized as a useful tool in the field of regenerative medicine and infertility. However, a small number of highly qualified SSCs are resided in the seminiferous tubule of testis, resulted in developing effective in-vitro culture system of SSCs for solving simultaneously quantitative and qualitative problems. Presently, SSCs can be enriched on testicular stromal cells (TSCs), but there are no systematic researches about TSC culture. Therefore, we tried to optimize culture condition of TSCs derived from mouse with different strains. For these, proliferation and viability were measured and compared by culturing ICR outbred or DBA/2 inbred mouse-derived TSCs at 35 or 37℃. In case of ICR strain, primary TSCs cultured at 37℃ showed significantly higher proliferation and viability than those at 35℃ and significant increase of proliferation and viability in sub-passaged TSCs was detected in the 35℃ culture condition. Moreover, sub-passage of primary TSCs at 35℃ induced no significant effects on proliferation and viability. In contrast, in case of DBA/2 strain, significantly improved proliferation were detected in the primary TSCs cultured at 35℃, which showed no significant difference in the viability, compared to those at 37℃. Furthermore, sub-passaged TSCs cultured at 37℃ showed no significant differences in proliferation and viability, compared to those at 35℃. However, with significant decrease of proliferation induced by sub-passage of primary TSCs at 35℃, no significant effects on proliferation and viability were resulted from sub-passage of primary TSCs at 37℃. From these results, culture temperature of primary TSCs derived from outbred and inbred strain of mouse could be separately optimized in primary culture and subculture.

The α-proteobacterium Wolbachia pipientis infects a wide range of arthropods and filarial nematodes. Wolbachia is maternally inherited and is known to induce reproductive anomalies such as cytoplasmic incompatibility, feminization, male killing and parthenogenesis induction (PI). Trichogramma kaykai is a tiny wasp that parasitizes on lepidopteran eggs. When a female of the wasp is infected with PI-Wolbachia, the female produces female offspring via gamete duplication without the aid of sperm. As she ages, however, the fraction of male offspring increases. In this study, we investigated the effect of host genetic background on the expression of sex ratio between isofemale lines. Virgin females of six isofemale lines were allowed to lay eggs individually for 10 days. There was the positive relationship between female age and the offspring sex ratio. Furthermore, the sex ratio was significantly different among isofemale lines, implying that the host genetic background had an effect on the sex ratio. Based on the results, evolution of symbiosis in terms of sex ratio and future experimental design are discussed.

The paternal sex ratio (PSR) chromosome is considered as an extremely selfish genetic element. It has only been found in the two hymenopteran insects- Nasonia and Trichogramma- with haplodiploid sex determination. When an egg is fertilized by sperm bearing PSR, the paternal genome is destroyed by PSR soon after fertilization resulting in haploid restoration and the egg develops into a male with only the maternal genome and PSR itself. Recently PSR is paid much attention, since it may be used for controlling haplodiploid pests. PSR can be successfully transferred from its natural host, T. kaykai to the novel host, T. deion. In the two hosts another sex ratio distorter, Wolbachia, is found. Wolbachia is a cytoplasmically inherited bacterium that induces parthenogenesis in this genus resulting in female offspring production without fertilization. The transmission efficiency of PSR in T. deion is lower than that in T. kaykai and is negatively influenced by the Wolbachia infection. The results show that 1) there is a negative host genetic background effect on the transmission of PSR in the novel host, 2) the transmission efficiency becomes even lower, when PSR males are infected with Wolbachia. The results imply that complex interactions among the bacterium, PSR and the species specific genetic background.

Near-isogenic lines (NILs) carrying bacterial blight resistance genes (Xa4, xa5 and Xa21) were developed in japonica rice using Suweon345 as genetic background. NILs were selected by gene specific DNA markers and inoculation of K1 or K3a race. NILs conferring Xa4 were resistant to K1, K2, K3, and moderately resistant to K3a. NILs conferring xa5 were resistant to K1, K2, K3, and K3a. NILs having Xa21 were susceptible to K1, while resistant to K2, K3 and K3a. Target genes of NILs with the genetic background of Suweon345 were also confirmed by using eleven Philippines races and International Rice Bacterial Blight (IRBB) NILs carrying Xa4, xa5 and Xa21. All NILs had no significant difference from their recurrent parents in the major agronomic traits except for panicle length and brown rice 1,000 grain weight. Heading date of NILs ranged from Aug. 10 to Aug. 11, which was similar to that of recurrent parent, Suweon345. Culm length, number of grains per panicle and ratio of ripened grain of NILs were similar to those of Suweon345. Milled rice of NILs was ranged from 4.82 to 4.93MT/ha. These NILs will be useful for improving resistance to K3a race of bacterial blight pathogens in Korean japonica cultivars.

최근 비교적 재배면적이 넓은 일미벼, 대산벼, 동안벼와 이들의 모본들로 구성된 23품종의 유전적 다양성을 조사한 결과를 요약하면 다음과 같다. 1. 1999년에서 2001년에 잎도열병과 이삭도열병이 심하게 발병한 일미벼. 대산벼, 동안벼는 밀양95호를 모본으로 육성되었다. 2. 밀양95호를 모본으로 하는 품종들에 대해 SSR 마커를 이용하여 유전적 다양성을 조사한 결과 57개 유전자좌위에서 170개(평균 3.0개)의 alleles이 관찰되었으며 대립유전자수는 2개에서 7개까지 다양하였다. 특히, RM249, RM206, OSR20은 6개 이상의 대립유전자를 가져 근연의 자포니카 벼품종간의 유전적 다양성 평가에 유용하게 이용될 수 있을 것으로 판단된다. 3.증폭된 밴드의 유무를 바탕으로 품종간 유전적 거리를 산출하여 군집분석을 실시한 결과 크게 4개의 군으로 나뉘었으며 일미벼, 대산벼, 동안벼는 모본인 밀양95호와 유전적 배경이 매우 유사하였다. 4. 같은 계보상의 품종들에 대해 도열병 저항성 유전자 인근의 마커를 이용한 유전적 다양성 분석결과가 계보도와 일치하여 이들 품종들의 저항성 유전자형 또한 유사할 것으로 추청할 수 있었다. 5. 조사품종의 계보상의 allele 전이를 분석한 결과 11번 염색체의 RM254의 3번째 allele과 12번 염색체의 OSR32의 2번째 allele이 밀양95호로부터 계속 전이되었음을 알 수 있었으며 이들 alleles의 도열병 이병화와의 연관성 여부는 추후에 검토가 이루어져야할 것이다.