Lee, Jang-Wook
(Genetics and Breeding Research Center, NFRDI)
,
Lee, Young Mee
(Genetics and Breeding Research Center, NFRDI)
,
Lee, Jeong-Ho
(Genetics and Breeding Research Center, NFRDI)
,
Noh, Jae Koo
(Genetics and Breeding Research Center, NFRDI)
,
Kim, Hyun Chul
(Genetics and Breeding Research Center, NFRDI)
,
Park, Choul-Ji
(Genetics and Breeding Research Center, NFRDI)
,
Park, Jong-Won
(Genetics and Breeding Research Center, NFRDI)
,
Hwang, In Joon
(Genetics and Breeding Research Center, NFRDI)
,
Kim, Sung Yeon
(Genetics and Breeding Research Center, NFRDI)
Fish larvae are immediately exposed to microbes from hatching to maturation of their lymphoid organs, therefore effective innate mechanisms is very important for survival in such an environment. The key component of innate immune system, C3 is central protein of all activation pathways of the comple...
Fish larvae are immediately exposed to microbes from hatching to maturation of their lymphoid organs, therefore effective innate mechanisms is very important for survival in such an environment. The key component of innate immune system, C3 is central protein of all activation pathways of the complement system, leading to inflammatory reactions, such as opsonisation, chemotaxis, and cell lysis of pathogens. Although, innate mechanisms is essential for survival in the early stage of development, little is known about defence mechanisms. In this study, the alignment analysis showed that amino acid sequence of C3 from olive flounder liver EST homologous to other known C3 sequences with 73-99% identity. Also, we examined the tissue distribution of olive flounder C3 and analyzed expression pattern from the fertilized egg until 28 days post hatching. As a result, olive flounder C3 mRNA was expressed only in the liver and the mRNA level more increased as developmental proceed during the early stage. These results may suggest that olive flounder C3 plays an important function in the early immune response of olive flounder larvae.
Fish larvae are immediately exposed to microbes from hatching to maturation of their lymphoid organs, therefore effective innate mechanisms is very important for survival in such an environment. The key component of innate immune system, C3 is central protein of all activation pathways of the complement system, leading to inflammatory reactions, such as opsonisation, chemotaxis, and cell lysis of pathogens. Although, innate mechanisms is essential for survival in the early stage of development, little is known about defence mechanisms. In this study, the alignment analysis showed that amino acid sequence of C3 from olive flounder liver EST homologous to other known C3 sequences with 73-99% identity. Also, we examined the tissue distribution of olive flounder C3 and analyzed expression pattern from the fertilized egg until 28 days post hatching. As a result, olive flounder C3 mRNA was expressed only in the liver and the mRNA level more increased as developmental proceed during the early stage. These results may suggest that olive flounder C3 plays an important function in the early immune response of olive flounder larvae.
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제안 방법
In order to investigate of olive flounder C3 mRNA expression during early development, we analyzed the temporal expression pattern of olive flounder C3 at the four embryos stages and ten larval stages (Fig. 3). As a result, the mRNA level of olive flounder C3 was not detected from fertilized egg (F) to blastula (E2).
The homologs search of the EST sequence with the corresponding amino acid sequence was examined using BLASTX search of GenBank and sequence alignment was performed with the ClustalW program (http://www.ch.emb net.org/software/ClustalW. html).
To investigate the tissue specific expression of the complement component C3 gene, oligonucleotide specific primers were designed according to previously reported C3 gene in olive flounder (GenBank accession no. AB021653), and then we examined by RT-PCR using isolated various tissues from 2-year-old healthy olive flounder.
데이터처리
The data was statistically analized by one-way ANOVA after arcsine transformation when needed, and followed by Duncan’s multiple-range test.
이론/모형
0×e－4 were considered to be significant. Multiple alignments of the amino acid were constructured using the ClustalW program using default parameters (Thompson, 1994) and visualized with MEGA 3.1 (Kumar et al., 2004).
The ratio between the housekeeping gene content in standard samples (106 copies) and test samples were defined as the normalization factor. The fold changes of C3 mRNA in different developmental stages or tissues were calculated using standard deltadelta-Ct method (Pfaffl, 2001).
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