Vector-borne diseases are transmitted to humans by blood-feeding arthropods such as mosquitoes, ticks, and fleas. These cold-blooded animals are influenced by environmental change. A recent report by IPCC showed that the emission of greenhouse gases has already changed world climates. Heat waves in Europe, rises in global mean sea level, summer droughts and wild fires, more intense precipitation, and increasing numbers of large cyclones, hurricanes and typhoon may be typical example of extreme climate phenomena related to global warming. High temperatures during winter season may increase survival rate among arthropod vectors in Temperate Zone. Warming may accelerate the spread of mosquitoes such as Aedes albopictus in the northern parts of Japan and European countries. The spread of the mosquito vector through global used-tire trading in recent decades to Africa, the Mideast, Europe, and North and South America caused an outbreak of Chikungunya fever in north Italy in 2007.

Thailand suffers from one of the highest rates of dengue and dengue haemorrhagic fever in the world. Since there is no vaccine or any specific treatments for dengue, currently, the only effective way to prevent dengue fever and dengue hemorrhagic fever is vector control. The knowledge of basic biology, ecology, behavior and survival factors of Ae. aegypti is crucial to control mosquitoes. The impact of the environmental factors; temperature, rainfall, and humidity, on the ecology, biology, development, vector density, behavior, fitness of mosquito vectors, and the mosquito-borne disease transmission dynamics will be presented and discussed.

Ecological changes, both man-made and natural, have changed the landscape of the Republic of Korea following the end of Japanese occupation at the end of World War II (1939-1945). During the Japanese occupation, forested hillsides were cleared and wood products shipped to Japan, leaving the hills and mountains largely covered by grasses and other shrub vegetation. Following WWII, the country of Korea was divided into North and South Koreas, with North Korea under communist powers, while South Korea was established as a democratic government. In South Korea poverty was rampant and local populations scavenged for wood for cooking and heating during the cold winters. Just as economic conditions were increasing, South Korea was attached by North Korea, beginning a long drawn-out conflict from 1950 to the summer of 1953, whereby an armistice was signed and an often uneasy peace between the two countries continues today. Again, the Republic of Korea emerged from a War as an impoverished country with treeless country sides, hills, and mountains. In the 1960’s, president Chung-Hee Park established a tree planting policy to reestablish long-ago lavish forested mountains and hillsides that make up more than 70% of the South Korean landscape. Today, mountains and hillsides are generally not used for agriculture and are completely forested, with planted groves and volunteer trees ranging in age from 10-50 years. These forested areas have led to increased protection for large and small mammals and birds, increasing the potential for zoonotic pathogens that there ectoparasites harbor to be transmitted to man during work and leisure activities. While forested areas provided an expanded habitat for some animals, agriculture expanded and modernized, resulting in short-cut grasses on banks separating much of the rice paddies, orchards, ditches, and dry-land farming that increased competition for small mammal habitat. As a result of increased surveillance of small and large animals and their ectoparasites, the increased prevalence of known pathogens and identification of new pathogens, especially those harbored by ticks, has demonstrated the presence of tick-borne encephalitis, several species of spotted fever group Rickettsia, and a host of other zoonotic diseases in wild animals, their ectoparasites, and man. As tick-borne diseases are not reportable diseases in Korea, the extent and impact on civilian and military populations is unknown as diagnoses are often likely sought for the wrong pathogen. While agriculture modernized, military training sites largely consist of unmanaged lands with tall grasses that are conducive to large and small mammal populations, which are host to a number of zoonotic diseases, e.g., hantaviruses, scrub typhus, murine typhus, leptospirosis, tick-borne encephalitis, spotted fever group rickettsial pathogens, Lyme disease, bartonellosis, etc. that impact on military populations training in those areas. The impact of training sites habitat modification has not been assessed, but for many sites is impractical. To reduce health risks of vectorborne diseases, the US Army has established better housing (tents to barracks with screened windows and air conditioning) at some field training sites, reducing the potential for the transmission of mosquito-borne pathogens (malaria and Japanese encephalitis virus). The increased use of permethrin-treated all climate uniforms (ACUs) and repellents, also reduce the potential for transmission of mosquito-, tick-, mite-, and flea-borne pathogens. However, training conditions at some field training sites remain largely unchanged, with personnel working and sleeping in tents that abut to forested areas where animals and their ectoparasites are present. While some training and maneuver sites are well planned for weapons qualifications of wheeled and tracked vehicles, others sites are in areas of unmanaged lands which are rodent infested. Increased surveillance by the 65th Medical Brigade not only provides a baseline and relative distribution for vector-borne diseases in Korea, but also provides disease trends and risk assessments that are necessary for protecting US military personnel training in Korea.

The sweet potato whitefly Bemisia tabasi is one of the most important pests of various horticultural crops. In addition, B. tabaci is a vector of many plant-pathogenic viruses and cause a serious secondary damage to crop plants. Association of plant-pathogenic virus with vector insects is known to be effective on the transmission capacity, fecundity, longevity of vectors including whiteflies. However, the interactive mechanisms between virus and vector insects are still poorly understood. Recently, a serious damage caused by virus disease together with B. tabasi emergence was identified at tomato glasshouse in Tongyoung. We detected the signals of Tomato Yellow Leaf Curl Virus (TYLCV) in tomato leaves and vector whiteflies using PCR amplification and confirmed its presence by those sequence comparison. To determine the effects of TYLCV acquisition on physiological status of vector whiteflies, transcript levels of genes that associated with metamorphosis, metabolism, stress and immune processes were compared between TYLCVinfected whiteflies and non-infected ones. Generally, the transcript levels of virus-infected whiteflies were lower than those of non-infected ones. In addition, the associations of endosymbiont levels within whiteflies were discussed in aspect of the acquisition and transmission of TYLCV.

Bacillus thuringiensis 1-3 (Bt 1-3), isolated from Korean soil sample, showed high insecticidal activity against Plutella xylostella. Recently, we improved plasmid capture system donor-s (PCS-S) by inserting attB sites including lacZ between transposable elements (designated as pTroy), to reduce background and construct E. coli-Bt shuttle vector. Through in vitro transposition with total plasmid DNA of Bt 1-3, at least 6 different size plasmids of Bt 1-3 were cloned. Among them, 47 clones which have approximately 10 kb plasmid in size were sequenced and 5 contigs were assembled. These contigs showed partial similarity with two known plasmids, pGI3 or pBMB175, separately. These cloned plasmids will acquire erythromycin resistance by BP recombination reaction with pDonrattPEm vector. After transformation into Bt cells, final erythromycin resistant Bt cell might contain novel E. coli-Bt shuttle vector. This scheme proposes that pTroy and pDonr-attPEm system can easily construct new shuttle vector by in vitro transposition, BP reaction, and erythromycin selection with any Bt plasmids.

본 연구는 돼지 B-casein 유전자 위치에서 EGFP가 발현될 수 있는 knock-in 벡터를 구축하기 위하여 실시되었다. 돼지의 B-casein 유전자를 이용하여 knock-in 벡터를 구축하기 위해 돼지의 태아 섬유아세포로부터 B-casein 유전자를 동정하였고 EGFP, SV4O polyA signal을 동정하였다. Knock-in 벡터는 5' 상동 영역 약 5 kb와 3' 상동 영역 약 2.7 kb로 구성되어있으며, positive selection marker로 neor 유전자를, negative selection marker로 DT-A 유전자를 사용하였다. 구축된 knock-in 벡터로부터 EGFP의 발현을 확인하기 위하여 생쥐 유선 세포인 HC11 세포에 knock-in 벡터를 도입하였다. 그 결과 EGFP의 발현을 HC11 세포에서 확인하였다. 이와 같은 결과로서 이 block-in 벡터는 knock-in 형질전환 돼지를 생산하는데 사용될 수 있을 것으로 생각된다.

Niemann-Pick type C disease (NPC), known an autosomal recessive disorder, is characterized by abnormal accumulation of cholesterol and glycolipid in late endosome/lysosome. The 95% case in NPC patients are caused by mutation of NPC1 gene, whose gene product is 13 transmembrane-domains and a sterol-sensing domain. The compensation of NPC1 by transgene expression in NPC1-deficient mouse recovered life span and sterility, but neurological correction incompletely, suggesting a possibility of gene therapy. Thus, in order to develop virial expression system for gene therapy of NPC, we isolated NPC1 cDNA and constructed Sindbis viral expression system for verification. NPC1 expression by Sindbis viral expression reduced the accumulation of cholesterol induced by treatment of U18666A and progesterone in both baby hamster kidney (BHK) cells and cultured hippocampal neurons. In addition, NPC1 expression increased the immunoreactivities of post-synaptic density protein 95kDa (PSD-95) in dendrites. On the basis of these results, we constructed a lentiviral vector for NPC1 expression and examined its expression in hippocampal cultured neurons.

본 연구에서는 방향벡터(direction vector)를 이용한 지역 탐색법과 유전자 알고리즘을 결합한 새로운 알고리즘인 D-GA를 제안한다. 새로운 개체(individual)를 찾기 위한 방향벡터로는 진화과정 중에 습득되는 정보를 활용하기 위한 학습방향벡터(Loaming direction vector)와 진화와는 무관하게 한 개체의 주변을 탐색하는 랜덤방향벡터(random direction vector) 등 두 가지를 구성하였다. 그리고, 10 부재 트러스 설계 문제에 단순 유전자 알고리즘과 D-GA를 적용하여 최적화를 수행하였고, 그 결과를 비교 검토함으로써 단순 GA에 비하여 D-GA의 정확성 및 효율성이 향상되었음을 확인하였다.

본 연구에서는 외래 유전자의 지속적인 발현에 의한 형질 전환 개체나 세포의 생리적인 부작용을 최소화하기 위하여 hTPO 유전자의 발현을 조절할 수 있는 tetracycline-inducible retrovirus vector system을 구축하고자 하였다. hTPO 유전자는 사람의 간암세포인 HepG2에서 분리한 RNA를 주형으로 하여 RT-PCR 방법을 이용하여 확보하였으며, 이 유전자를 MLV 유래의 vector에 도입하여 pLNChTPOW를 재조합하였다. 재조합한 vector는 GP2 293 포장세포에 도입하여 바이러스를 생산하였으며, 이 바이러스를 이용하여 감염시킨 여러 표적세포에서 hTPO의 발현을 확인하였다. 또한, hTPO의 발현을 유도적으로 조절할 수 있도록 하기 위하여 hTPO를 one vector 형태의 Tet-On vector system에 도입하였으며, 발현의 유도 조건에서 보다 강한 발현을 위하여 WPRE 서열을 여러 위치에 도입하였다. 구축한 Tet system의 발현 조절 정도는 각 바이러스를 감염시켜서 구축한 CEF와 PFF 세포에서 RT-PCR과 Western blot, 그리고 ELISA 방법을 이용하여 확인하였다. 그 결과, CEF에서는 WPRE 서열이 hTPO 유전자의 3'에 위치한 경우에서, PFF에서는 WPRE가 rtTA의 3'에 위치한 경우의 vector system에서 가장 높은 발현율과 유도율을 나타내었다. 이는 Tet system에서의 hTPO 유전자 발현 조절이 매우 효율적으로 이루어지며, 세포주에 따른 의존적인 조절 양상을 보이는 것을 의미한다. 따라서 hTPO의 대량 생산을 위한 생체 반응기로서의 형질 전환 동물의 개발을 보다 효율적으로 수행하려면 적절한 Tet system이 선별적으로 적용되어야 할 것이다.