为了解慢性密度胁迫下绿鳍马面鲀(Thamnaconus septentrionalis)肌肉组织的分子响应机制,挖掘密度胁迫下的相关信号通路和差异表达基因,设置2个养殖密度组(即100尾/m^(3)的中密度组和500尾/m^(3)的高密度组)对绿鳍马面鲀肌肉转录组进行...为了解慢性密度胁迫下绿鳍马面鲀(Thamnaconus septentrionalis)肌肉组织的分子响应机制,挖掘密度胁迫下的相关信号通路和差异表达基因,设置2个养殖密度组(即100尾/m^(3)的中密度组和500尾/m^(3)的高密度组)对绿鳍马面鲀肌肉转录组进行了研究。分别于第25天和第50天时获得养殖绿鳍马面鲀的肌肉组织,利用Illumina测序平台对肌肉组织样本进行了转录组测序,得到103.3 Gb高质量测序数据。对照开始时暂养的绿鳍马面鲀,各试验组均筛选出大量的差异表达基因。通过GO(Gene Ontology)功能注释与KEGG(Kyoto Encyclopedia of Genes and Genomes)功能富集分析,筛选出涉及破骨细胞分化、MAPK、PI3K-Akt等路径以及黏着斑相关的信号传导机制,发现nfatc1、tgfbr2、map3k14、pparg、fos、flna、flnc、fn1、tln1、thbs1等关键基因在绿鳍马面鲀肌肉组织中的差异性表达。研究表明,这些差异表达基因可能在调节绿鳍马面鲀的细胞生长和分化、免疫能力等方面发挥关键作用。展开更多
There are 10 percent to 15 percent genes expression in certain cells during the life time of fishes like other vertebrates. The genes were different at different development stage,under different physiological conditi...There are 10 percent to 15 percent genes expression in certain cells during the life time of fishes like other vertebrates. The genes were different at different development stage,under different physiological conditions, and in different kinds of cells. So comparing the differences of gene expression in different cells can help us understand the genetic nature of phenotypic differences, and understand the basic information of life period, and find the genes in relation to development and diseases, and finally benefit mankind. Several methods were developed to clone differential expression gene in recent years. They are subtractive hybridization (SH), differential display (DD), representional difference analysis (RDA), and so on. These methods all have postive influences on cloning special genes, but they all have some defects, such as higher false-positive, lower replication, lower sensitive and difficulty to manipulate. Suppression subtractive hybridization (SSH) was developed by Diatchenko et al in 1996. SSH was based on suppression PCR and combines normalization and subtraction in a single procedure. It is a more effective and convenient method than all others mentioned above. The principle and the rules of manipulation of SSH in detail was illuminated and the novel genes cloned by SSH was listed. They are immune related genes and reproduction and development related genes. The reproduction and development related genes are as follows: ZP3, Cyclin A2, CB102, YA2, FSTRAP. The immune related genes are as follows: NKEF(natural killer enhancing factor),CC chemokine, CXCR1, CXCR2, CXCR4, AIF-1(allograft inflammatory factor-1), IL-1β(inteleukin-1), FcεRIγ (γ submit of high affinity Fc receptor for IgE), SSA(serum amyloid A), LECT2(leucocyte cell-derived hemotaxin 2), GMFβ(glia maturation factorβ), CD45, Lysozyme C, PBEF (Pre-B cell enhancing factor), C-type lectin, PTX(Pentraxin), IL-1RII, IL-8-like CXC chemokine, TF(tissue factor), trout chemokine 2, TNF decoy receptor, M17. Some subtracted cDNA libraries were also built by SSH method.展开更多
文摘为了解慢性密度胁迫下绿鳍马面鲀(Thamnaconus septentrionalis)肌肉组织的分子响应机制,挖掘密度胁迫下的相关信号通路和差异表达基因,设置2个养殖密度组(即100尾/m^(3)的中密度组和500尾/m^(3)的高密度组)对绿鳍马面鲀肌肉转录组进行了研究。分别于第25天和第50天时获得养殖绿鳍马面鲀的肌肉组织,利用Illumina测序平台对肌肉组织样本进行了转录组测序,得到103.3 Gb高质量测序数据。对照开始时暂养的绿鳍马面鲀,各试验组均筛选出大量的差异表达基因。通过GO(Gene Ontology)功能注释与KEGG(Kyoto Encyclopedia of Genes and Genomes)功能富集分析,筛选出涉及破骨细胞分化、MAPK、PI3K-Akt等路径以及黏着斑相关的信号传导机制,发现nfatc1、tgfbr2、map3k14、pparg、fos、flna、flnc、fn1、tln1、thbs1等关键基因在绿鳍马面鲀肌肉组织中的差异性表达。研究表明,这些差异表达基因可能在调节绿鳍马面鲀的细胞生长和分化、免疫能力等方面发挥关键作用。
文摘There are 10 percent to 15 percent genes expression in certain cells during the life time of fishes like other vertebrates. The genes were different at different development stage,under different physiological conditions, and in different kinds of cells. So comparing the differences of gene expression in different cells can help us understand the genetic nature of phenotypic differences, and understand the basic information of life period, and find the genes in relation to development and diseases, and finally benefit mankind. Several methods were developed to clone differential expression gene in recent years. They are subtractive hybridization (SH), differential display (DD), representional difference analysis (RDA), and so on. These methods all have postive influences on cloning special genes, but they all have some defects, such as higher false-positive, lower replication, lower sensitive and difficulty to manipulate. Suppression subtractive hybridization (SSH) was developed by Diatchenko et al in 1996. SSH was based on suppression PCR and combines normalization and subtraction in a single procedure. It is a more effective and convenient method than all others mentioned above. The principle and the rules of manipulation of SSH in detail was illuminated and the novel genes cloned by SSH was listed. They are immune related genes and reproduction and development related genes. The reproduction and development related genes are as follows: ZP3, Cyclin A2, CB102, YA2, FSTRAP. The immune related genes are as follows: NKEF(natural killer enhancing factor),CC chemokine, CXCR1, CXCR2, CXCR4, AIF-1(allograft inflammatory factor-1), IL-1β(inteleukin-1), FcεRIγ (γ submit of high affinity Fc receptor for IgE), SSA(serum amyloid A), LECT2(leucocyte cell-derived hemotaxin 2), GMFβ(glia maturation factorβ), CD45, Lysozyme C, PBEF (Pre-B cell enhancing factor), C-type lectin, PTX(Pentraxin), IL-1RII, IL-8-like CXC chemokine, TF(tissue factor), trout chemokine 2, TNF decoy receptor, M17. Some subtracted cDNA libraries were also built by SSH method.
