Plant-parasitic nematodes are the most devastating group of plant pathogens worldwide and are extremely challenging to control. In the present study, we have performed a genome wide analysis to identify common genes among four nematode species consisting of root-knot nematodes (Meloidogyne incognita and Meloidogyne hapla), cyst nematode (Heterodera glycines), and free living nematode (Caenorhabditis elegans) respectively. Using their whole genome sequences, we predicted 15,274 genes from M. incognita, 38,149 genes from M. hapla, 8,061 genes from H. glycines and 23,894 genes from C. elegans, where, among the predicted genes, 1,358, 1,350, 1,401, 1,365 respectively from each nematode, code for common groups of proteins. Further, 2,067, 2,086, 1,566, 2,903 genes were recollected using Clusters of Orthologous Groups (COG) database. Under our search criteria, a total of 800 common genes were identified in all the four studied nematode genomes. The most annotated conserved genes were obtained from four different species using Basic Local Alignment Searching Tool (BLAST). Uni- Prot Taxon identifier database was used to elucidate their taxonomic classification such as 698 genes under kingdom Metazoa, 660 genes confined to Nematoda, 290 genes in Chordata and 660 genes falling under class Chromadorea. The biochemical characterization of proteins expressed by these genes was examined using Pedant-Pro sequence analysis. The protein length, molecular weight, isoelectric point (pI), and transmembrane domain of the coded proteins were at a range of 300 to 999 amino acids (40.9%), molecular weight of over 100 kDa (96%), pI from 4.5 to 5.5 (27.6%) and 0 (56.6%), respectively. To classify protein function, the obtained BLAST hits were assigned to Gene Ontology classification scheme. The fractions of protein function were distributed as cellular component, biological processes and molecular function of the cell (22.2%), multicellular organism process (15.8%) and binding (48.3%), respectively. The current study provides an excellent resource for nematode functional genomics studies, which can be utilized further for studies on role of genes involved in nematode biological processes.

Human tissue-type plasminogen activator (t-PA) is responsible for fibrin-specific plasminogen activation and plays a key role in fibrinolysis thereby aiding breakdown of blood clots in the vasculature. In the present study, in order to develop a system for production of recombinant st-PA and t- PAHis6 proteins in transgenic rice seeds, a DNA fragment encoding t-PA gene was selected and cloned to a plant binary vector (pMJ21) harboring a rice GluB1 promoter, an N-terminal signal peptide of the rice glutelin B1 protein and a Pin II terminator. The constructed plasmid was transformed into Agrobacterium tumefaciens LBA4404 (pSB1) to facilitate introduction into rice callus. The insertion of the st-PA and t-PAHis6 genes into the genome of transgenic rice seeds and their transcripts were confirmed using PCR, and Southern blot as well as RT-PCR, respectively. The highest level of recombinant st-PA expression as determined by enzyme-linked immunosorbent assay (ELISA) was found to be 2,916 ng/total soluble protein (mg) in transgenic rice seeds. The amount of recombinant proteins expressed in transgenic plants was estimated to range from 634 ~ 2,916 ng/TSP mg (st-PA) and 925 ~ 2,640 ng/TSP mg(t- PAHis6), respectively. Immuno-blot analysis of transgenic rice seeds revealed single bands of approximately 68-kDa representing recombinant st-PA and t-PAHis6 proteins. These results demonstrate the expression and in vivo activity of recombinant st-PA and t-PAHis6 in transgenic rice seeds. This study is a promising endeavor for production of recombinant pharmaceutical proteins using rice seed system.

본 연구는 새로운 항산화 및 미백 기능성 화장품 성분을 개발하기 위해 기존의 항산화와 미백 기능성 성분으로 알려진 두 가지 성분을 혼합한 새로운 조성을 개발하고, 이를 이용해 항산화 및 미백효과를 알아보고자 하였다. 두 가지 성분은 항산화와 멜라닌 생성억제 기능이 알려진 sulforaphane과 멜라닌 생성 억제 효과가 알려진 PF-3758309이다. 각 성분 및 혼합 조성에 대한 세포독성을 조사하여 세포독성이 나타나지 않는 농도를 결정하였다. 단독 처리 시 sulforaphane는 10 μ M까지, PF-3758309는 100 nM까지 세포독성이 없다는 것을 확인하였다. 두 화합물 10 μ M sulforaphane과 1 nM PF-3758309를 병합 처리 시 세포독성은 없었다. LPS 자극에 의해 생성된 일산화질소는 sulforaphane과 PF-3758309의 병합 처리시 현저히 감소되었다. α-MSH 에 의한 멜라닌 생성 역시 병합 처리에 의해 억제되었다. 화장품 소재로 사용 가능성을 확인하기 위해 혼합 조성 시료에 대한 인체적용시험을 실시하였다. 혼합조성 시료는 피부첩포에 의한 안전성 평가는 무자극으로 판정되었고, 피부밝기 증가효과와 피부 멜라닌 침착을 억제함을 확인하였다. 이러한 결과들로 혼합 조성물이 항산화와 미백 기능이 있는 새로운 기능성 화장품 조성으로 개발될 가능성을 확인할 수 있었다.