Room temperature 1.55 mu m emission from InAs quantum dots grown on (001)InP substrate by molecular beam epitaxy

Self-assembled InAs nanostructures on (0 0 1)InP substrate have been grown by molecular beam epitaxy (MBE) and evaluated by transmission electron microscopy (TEM) and photoluminescence (PL). It is found that the morphologies of InAs nanostructures depend strongly on the underlying alloy. Through introducing a lattice-matched underlying InAlGaAs layer on InAlAs buffer layer, the InAs quantum dots (QDs) can be much more uniform in size and great improvement in PL properties can be attained at the same time. In particular, 1.55 mu m luminescence at room temperature (RT) can be realized in InAs QDs deposited on (0 0 1)InP substrate with underlying InAlGaAs layer. (C) 2000 Published by Elsevier Science B.V. All rights reserved.

灵长类认知相关基因-Microcephalin和PACAP 前体基因的分子进化研究

人是生命演化的产物。然而由于具有一个超级大的大脑和高级的认知能力， 人类在自然界中又显得十分与众不同。在灵长类进化过程中，脑容量大小和认知 能力高低可以明显的分成几个等级，由高到低依次为人类、大猿和其它灵长类。 现在一般认为，人类超级大的大脑的形成是适应性选择(达尔文正选择)的结果。 但人类高级认知的起源一直还是个未解之谜。随着分子进化理论和比较基因组学 的发展，人们有可能从分子进化的角度去探讨高级认知的起源问题，并揭示其遗 传学机制。 本文从两个不同的角度入手，通过对认知相关的候选基因的分子进化研究来 探索高级认知起源的遗传学机制。1. 从表型出发，发现microcephalin 基因跟小 脑症相关。我们推测，该基因可能在人类起源过程中发生一些突变，导致脑容量 增大。2. 比较基因组学方法发现垂体腺苷酸环化酶激活肽(PACAP)前体基因在人 类支系中存在正选择。这类受正选择的神经相关基因可能在进化过程中对认知能 力的提高发挥重要作用。 Microcephalin 基因在人脑的发育过程中起重要作用。导致该基因蛋白质翻译 提前终止的突变，会引起人类小脑症，患者脑容量回复到早期原始人的大小。我 们对人群和12 个非人灵长类物种的microcephalin 基因编码区进行测序。所测的 非人灵长类物种包括大猿、小猿、旧大陆猴和新大陆猴。结果显示，microcephalin 基因在人群中具有较高的多态性。在人群中编码区有22 个多态位点，其中有15 个是错义突变位点。中性检验结果显示，microcephalin 基因的高多态性是人类群 体扩张和达尔文正选择共同作用的结果。灵长类各支系的进化分析表明， microcephalin 基因在大猿的起源过程中受到较强的正选择信号。这正好和小猿向 大猿进化过程中大脑容量急剧增大，认知能力大幅度提高的事实相符。单个氨基 酸编码位点检验显示，microcephalin 基因有五个氨基酸位点在灵长类的进化过程中存在正选择信号。这几个位点可能在灵长类进化和人类起源过程中，对脑容量 的增大和认知能力的提高起到了重要的作用。 PACAP 是一个神经系统中高表达的神经肽，与神经发生和神经信号传导有 关。该基因的氨基酸序列在脊椎动物的进化过程中非常保守，表明PACAP 前体 基因在这一进化历程中受到强烈的功能限制。然而比较序列分析显示，自从人与 黑猩猩分歧以后，人类支系中PACAP 前体基因有明显的加速进化。由于存在强 烈的正选择作用，人类支系的氨基酸替换速率是其它哺乳动物支系的至少7 倍。 PACAP 前体基因中有7 个人类特异的氨基酸改变，而这些氨基酸在从鼠类到大 猿的所有物种中都是保守的。蛋白质结构分析结果显示，人类起源过程中，PACAP 前体基因可能编码一个新的神经肽，并在人脑中具有功能。因此，在人类起源过 程中，PACAP 前体基因发生了适应性的改变，可能对人类认知能力的产生起到 了重要作用。这个新神经肽是否存在及其功能还有待于进一步功能实验的验证。 总之，本文通过分子进化分析，发现了两个在灵长类进化过程中对认知能力 的提高起关键作用的基因(Microcephalin 基因、PACAP 前体基因)。Microcephalin 基因，在1400-1800 万年前，从小猿向大猿的进化过程中，可能对脑容量的增大 和认知能力的提高发挥了重要的作用。PACAP 前体基因则在最近的几百万年内， 现代人的起源过程中，对人脑的形成以及人类高级认知能力的产生可能发挥了重 要的作用。

基于ITS 序列和matK 基因序列的中国药用石斛及其混伪品的分子鉴定

为了从分子水平对中国药用石斛及其混伪品进行鉴定，本文选取了核rDNA ITS 序列和叶绿体DNA 的matK 基因序列进行研究。采用改良的CTAB 法提取石斛的基因组DNA，PCR 产物直接测序法对17 种（共32 份）药用石斛的核糖体内转录间隔区ITS 全序列进行测定，克隆测序法对12 种（共22 份）药用石斛的叶绿体的matK 基因序列进行测定，运用BioEd it，MEGA4.0 等生物软件分析了石斛属植物的rDNA ITS 序列及叶绿体的matK 基因序列的特征，比较了石斛属间、种间、种内不同居群（品种）间的序列碱基差异及遗传距离，应用邻接法构建分子系统树。主要研究结果如下： （1）建立了17 种（共32 份）药用石斛rDNA ITS 区碱基全序列数据库，其中，ITS1 的长度为228~234 bp，GC 含量为45.7%~53.0%，变异位点167 个，占总位点67.34%，信息位点106 个，占总位点42.74%，ITS2 长度为241~247 bp，GC含量为44.8%~55.7%，变异位点165 个，占总位点66.27%，信息位点115 个，占总位点46.18%。 （2）建立了12 种（共22 份）药用石斛的叶绿体matK 基因全序列数据库，叶绿体matK 基因长1410 bp，变异位点51 个，信息位点11 个。除了存在碱基替换的遗传变异外，还存在碱基的插入和缺失。 （3）通过ITS 序列比较分析了各材料间的遗传距离和碱基差异，属间的遗传距离为0.295，石斛种间的平均遗传距离为0.142，碱基相差2~156 个，种内各居群间的平均遗传距离为0.002，碱基相差1~2 个。属间的遗传距离大于种间的遗传距离，种间的遗传距离大于种内不同居群（品种）间的遗传距离。 （4）根据分析石斛叶绿体的matK 基因序列得到，外类群（密花石豆兰）与石斛属间最小遗传距离为0.027，石斛种间的平均遗传距离为0.008，种间最大的遗传距离0.014， 最小的遗传距离为0.003，碱基相差8~20 个。种内不同居群（品种）遗传距离为0.001，相差1~5 个碱基。 （5）利用17 种石斛的全序列数据库及遗传分析软件，通过对待检种rDNA I TS区进行序列测定，成功地对10 个待检种进行了鉴定，并且在原植物开花后得到了验证。 （6）运用12 种石斛的matK 基因全序列数据库及遗传分析软件，成功地对4个待检种进行了鉴定，同样在原植物开花后得到了验证。 （7）本文利用石斛的核糖体内转录间隔区ITS 序列和叶绿体的matK 基因序列数据库分别构建了NJ 树，外类群与石斛属间石斛种间以及种内不同居群（品种）间均能在NJ 树中明显分化开来，二者构建的分子系统树一致，为石斛的分子鉴定提供了依据。 In order to identify Chinese Herba Dendrobii and its adulterant species on molecular level, we studied the sequences of rDNA ITS and chloroplast matK gene. Genomic DNA of Dendrobium was extracted using the modified cetyltrimethyl ammonium bromide (CTAB) method. The PCR products of the rDNA ITS sequences of Dendrobium (32 materia ls) were purified and then sequenced. The PCR products of chloroplast matK gene of Dendrobium (22 materia ls) were purified, cloned and then sequenced. The characteristic of the sequences and the genetic dista nce were compared between Bulbophyllum odoratissimum and Dendrobium, Dendrobium interspecies, and different populations. Phylogenetic trees were constructed using the NJ method by the biology softwares including BioEd it, MEGA4.0 etc. The ma in results as follows: (1) It was built up that the database of rDNA ITS sequences of 17 species of Herba Dendrobii (32 materia ls). The ITS1 was 228~234 bp, the GC content accounting for 45.7%~53.0%. Its variable sites were 167, accounting for 67.34%. The Parsim-Informative positions were 106, accounting for 42.74%. The ITS2 was 241~247 bp, the GC accounting for 44.8%~55.7%. The variable sites were 165, accounting for 66.27%. The Parsim-Informative positions were 115, accounting for 46.18%. (2) The database of the chloroplast matK gene sequences was built up, which contained 12 species of Herba Dendrobii (22 materia ls). The matK gene sequences were about 1410bp in length. There were 51 variable sites and 11 Parsim-Informative sites. And there were nucleotides insertions and deletions in some species , in addition to the nucleotides substitutions. (3) The rDNA ITS sequences were compared and analyzed by the biology softwares. The genetic dista nce between Bulbophyllum odoratissimum and Dendrobium was 0.295. The avera ge genetic dista nce was 0.142 between Dendrobium species, and there were 2~156 variable nucleotides. The avera ge genetic dista nce between different populations was 0.002, and there were 2~156 variable nucleotides. The genetic dista nce between Bulbophyllum odoratissimum and Dendrobium was greater tha n that of Denrobium interspecies. Meanwhile, the genetic dista nce between Denrobium species was also greater tha n that of different populations (variaties). (4) The characteristics of the chloroplast matK gene sequences were obtained after analyzing by the biology softwares. The minima l genetic dista nce was 0.027 between Bulbophyllum odoratissimum and Dendrobium . The ma xima l genetic dista nce was 0.014 between Dendrobium species, and there were 20 variable nucleotides. The minima l genetic dista nce between populations was 0.003, and there were 8 variable nucleotides.The genetic dista nce between populations was 0.001, and there were 1~5 variable nucleotides. (5) The molecular Phylogeny tree was constructed on the database of rDNA ITS the sequences of 17 species of Herba Dendrobii using the biology softwares. Then we authenticated 10 materia ls on molecular level. What’s more, they had been proved when these pla nts flowered. (6) The molecular Phylogeny tree was built up on the database of chloroplast matK gene sequences of 12 species of Herba Dendrobii with the biology softwares.Then 4 materia ls were authenticated on molecular level. Moreover, they had also been proved when these pla nts were in flower. (7) The Phylogenetic trees were separately constructed on the sequences of rDNA ITS and chloroplast matK gene B. odoratissimum and Dendrobium all could be distinguished on the Phylogenetic trees. Meanwhile, the Phylogenetic trees based on two groups of sequences were coincident. rDNA ITS and matK gene sequence could be used as molecular markers for authentication of Herba Dendrobii.

A new potential radiosensitizer- multi-walled carbon nanotubes modified by ammonium persulfate

Here we prepare carbon nanotubes modified with ammonium persulfate, very short carbon nanotubes with 50-100 nanometer length was obtained, and the higher P potential of 52 mV was detected, these supporting the successful modification. HeLa cells were irradiated with P rays via adding or absent above functionalized carbon nanotubes (f- WCNTs) into cell culture medium with different concentration and radiation dosage. Confocal microscopy images and fluorescence-labeled DNA detection verified the successfully pure multi-walled carbon nanotubes (p-WCNTs) and f-WCNTs penetrated into cells. Compared with pure radiation, by MTT test, f-WCNTs induced cell death markedly with about 8.7 times higher than former one under little dose of radiation; meanwhile, no obvious toxicity was observed both in p-WCNTs and f-WCNTs without of radiation exposure. We hypothesized that large amount of hydroxyl and carbonyl organs on the surface of very short f-WCNTs changed into free radicals result from radiations led cell damage. These implied that f-WCNTs could be regarded as a new radiosensitizer.

Shell effect and capture cross sections in the synthesis of superheavy nuclei

The shell effect is included in the improved isospin dependent quantum molecular dynamics model in which the shell correction energy of the system is calculated by using the deformed two-center shell model. A switch function is introduced to connect the shell correction energy of the projectile and the target with that of the compound nucleus during the dynamical fusion process. It is found that the calculated capture cross sections reproduce the experimental data quantitatively at the energy near the Coulomb barrier. The capture cross sections for reaction (35) (80) Br + (82) (208) Pb -> (117) (288) X are also calculated and discussed.

红毛菜发育过程及其生理基础

红毛菜（Bangia Lyngb.）属于红藻门，与紫菜属同属红毛菜科，其味道和营养都优于紫菜。目前红毛菜栽培产业已在我国福建莆田展开，但栽培技术还有待提高。海藻栽培技术的发展和成熟依赖于对其生长发育过程的认识。本研究针对红毛菜发育过程及相关光合生理展开，并初步探讨了一采自山西娘子关泉淡水红毛菜群体（FWB）的系统地位。 色素突变标记的壳孢子萌发特征表明最初两次分裂产生的4细胞决定了完整植株的形态建成。成熟植株，为雌雄异体。雌性生殖器果胞的标志性分化结构为原始受精丝，环境因子是促发原始受精丝发展的外部因素，其膨大程度随受精的延迟而增大。原孢子是主要的无性生殖孢子类型，在不良环境中，藻体也会形成内生孢子或休眠包囊，或者藻体断裂后重新形成完整的植株。 红毛菜的生长发育很大程度上受环境因子的控制。高温不利于配子体的发育，15-20 ºC比较适宜。红毛菜无性繁殖的最适温度-光照组合为20 ºC-8 h，有性繁殖为15 ºC-12h。 不同发育阶段，PSII实际光合效率（Y(II)）与细胞的健康状况以及光合器官完整性及其在细胞内的分布有关，而与细胞的类型关系不大。健康的假根细胞、已分化未成熟的精子以及果孢子细胞均具有很高的Y(II)。色素体由中间位变为围周位，中央大液泡（营养藻丝）和大小纤维囊泡（成熟孢子与精子）的产生，使得细胞Y(II)降低。刚放散的壳孢子Y(II)很低，说明在壳孢子由贝壳基质释放到自由水体过程，光合作用受到一定程度抑制；而2h后，Y(II)开始恢复，rbcL的转录水平非常高，为孢子的萌发储备物质和能量需求。 在失水和低盐胁迫下，藻体均维持较高的Y(II)。干出处理至藻体重量不再变化，复水后Y(II)可回复初始水平。海生红毛菜在100％淡水培养基中（约20ºC）培养7天后，部分雄性藻体依然活着。从而体现了红毛菜位居高潮带的生理优势。 FWB终生行无性繁殖，藻体形态与发生以及染色体数目（4条）与海生群体没有区别。而rbcL-rbcS Spacer序列显示，红毛菜海生群体（无性和有性）具有完全相同的序列，而FWB与它们有5bp差异，但是与欧洲、北美地区的淡水群体仅1bp不同，初步说明所有淡水红毛菜群体具有共同的原始起源。

Molecular cloning and expression of a Toll receptor gene homologue from Zhikong Scallop, Chlamys farreri

Toll-like receptors (TLRs) are an ancient family of pattern recognition receptors, which show homology with the Drosophila Toll protein and play key roles in detecting various non-self substances and then initiating and activating immune system. In this report, the full length of the first bivalve TLR (named as CfToll-1) is presented. CfToll-1 was originally identified as an EST (expressed sequence tag) fragment from a cDNA library of Zhikong scallop (Chlamys farreri). Its complete sequence was obtained by the construction of Genome Walker library and 5' RACE (rapid amplification of cDNA end) techniques. The full length cDNA of CfToll-1 consisted of 4308 nucleotides with a polyA tail, encoding a putative protein of 1198 amino acids with a 5' UTR (untranslated region) of 211 bp and a 3'UTR of 500 bp. The predicted amino acid sequence comprised an extracellular domain with a potential signal peptide, nineteen leucine-rich repeats (LRR), two LRR-C-terminal (LRRCT) motifs, and a LRR-N-terminal (LRRNT), followed by a transmembrane segment of 20 amino acids, and a cytoplasmic region of 138 amino acids containing the Toll/IL-1R domain (TIR). The deduced amino acid sequence of CfToll-1 was homologous to Drosophila melanogaster Tolls (DmTolls) with 23-35% similarity in the full length amino acids sequence and 30-54% in the TIR domain. Phylogenetic analysis of CfToll-1 with other known TLRs revealed that CfToll-1 was closely related to DmTolls. An analysis of the tissue-specific expression of the CfToll-1 gene by Real-time PCR showed that the transcripts were constitutively expressed in tissues of haemocyte, muscle, mantle, heart, gonad and gill. The temporal expressions of CfToll-1 in the mixed primary cultured haemocytes were observed after the haemocytes were treated with 1 mu g ml(-1) and 100 ng ml(-1) lipopolysaccharide (LPS), respectively. The expression of CfToll-1 was up-regulated and increased about 2-fold at 6 h with the treatment of 1 mu g ml(-1) LPS. The expression of CfToll-1 was down-regulated with the treatment of 100 ng ml(-1) LPS. The results indicated that the expression of CfToll-1 could be regulated by LPS, and this regulation was dose-dependent. (c) 2006 Elsevier Ltd. All rights reserved.

Molecular cloning, expression of a big defensin gene from bay scallop Argopecten irradians and the antimicrobial activity of its recombinant protein

Antimicrobial peptides are important components of the host innate immune responses by exerting broad-spectrum microbicidal activity against pathogenic microbes. The first mollusk big defensin (designated AiBD) cDNA was cloned from bay scallop Argopecten irradians by expressed sequence tag (EST) and rapid amplification of cDNA ends (RACE) techniques. The scallop AiBD consisted of 531 nucleotides with a canonical polyadenylation signal sequence AATAAA and a poly(A) tail, encoding a polypeptide of 122 amino acids. The high similarity of AiBD deduced amino acid sequence with big defensin from Tachypleus tridentatus and Branchiostoma belcheri tsingtaunese indicated that AiBD should be a member of big defensin family. The expression of AiBD in various tissues was measured by using Northern blotting analysis. mRNA transcripts of AiBD could be detected in haemocytes of unchallenged scallops. The temporal expression of AiBD in haemolymph after Vibrio anguilarum challenge was recorded by quantitative real time PCR. The relative expression level of AiBD in haemolymph was up-regulated evenly in the first 8 h, followed by a drastic increase, and increased 131.1-fold at 32 h post-injection. These results indicated that AiBD could be induced by bacterial challenge, and it should participate in the immune responses of A. irradians. Biological activity assay revealed that recombinant AiBD could inhibit the growth of both Gram-positive and Gram-negative bacteria, and also showed strong fungicidal activity towards the expression host. Recombinant expression of AiBD made it possible to further characterize its functions involved in immune responses, and also provided a potential therapeutic agent for disease control in aquaculture. (c) 2006 Elsevier Ltd. All rights reserved.

Molecular cloning and responsive expression to injury stimulus of a defender against cell death 1 (DAD1) gene from bay scallops Argopecten irradians

Apoptosis is an active process of cell death, which is an integral part of growth and development in multicellular organisms. The defender against cell death 1 (DAD1), the regulatory protein to inhibit the apoptosis process, was first cloned from the bay scallop Argopecten irradians by randomly sequencing a whole tissue cDNA library and rapid amplification of cDNA end (RACE). The full-length cDNA of the A. irradians DAD1 was 607 bp, consist of a 5'-terminal untranslated region (UTR) of 63 bp, a 3'-terminal UTR of 205 bp with a canonical polyadenylation signal sequence AATAAA and a poly (A) tail, and an open reading frame of 339 bp. The deduced amino acid sequence of the A. irradians DAD1 showed 75.5% identity to Araneus ventricosus, 74.5% to Drosophila melanogaster, and 73.6% to Homo sapiens, Sus scrofa, Mesocricetus auratus, Rattus norvegicus and Mus musculus. Excluding the Saccharomyces cerevisiae DAD1 homologue, all animal DAD1 including A. irradians DAD1 homologue formed a subgroup and all plant DAD1 proteins formed another subgroup in the phylogenetic analysis. The A. irradians DAD1 was expressed in all examined tissues including adductor muscle, mantle, gills, digestive gland, gonad and hemolymph, suggesting that A. irradians DAD1 is expressed in most body tissues. Furthermore, the mRNA expression levels of A. irradians DAD1 gene of hemolymph were particularly high after injury, suggesting that the gene is responsive to injury stimuli.

Molecular cloning, characterization and expression of a novel serine proteinase inhibitor gene in bay scallops (Argopecten irradians, Lamarck 1819)

Serine protease inhibitors, critical regulators of endogenous proteases, are found in all multicellular organisms and play crucial roles in host physiological and immunological effector mechanisms. The first mollusk serine proteinase inhibitor (designated AISPI) cDNA was obtained from the bay scallop Argopecten irradians by randomly sequencing a whole tissue cDNA library and rapid amplification of cDNA ends (RACE). The full-length cDNA of the scallop serine protease inhibitor was 1020 bp, consisting of a 5'-terminal untranslated region (UTR) of 39 bp, a 3'-terminal UTR of 147 bp with a canonical polyadenylation signal sequence AATAAA and a poly(A) tail, and an open reading frame of 834 bp. The AISPI cDNA encoded a polypeptide of 278 amino acids with a putative signal peptide of 22 amino acids and a mature protein of 256 amino acids. The deduced amino-acid sequence of AISPI contained six tandem and homologous domains similar to that of Kazal-type serine protease inhibitors, including the conserved sequence C-X(7)-C-X(6)-Y-X(3)-C-X(2,3)-C and six cysteine residues responsible for the formation of disulfide bridges, indicating that the AISPI protein from bay scallop should be a member of the Kazal-type serine protease inhibitor family. The temporal expression of AISPI was measured by semi-quantitative RT-PCR after injury or bacterial challenge. After the adductor muscle was wounded or injected with Vibrio anguillarum, the expression of AISPI mRNA in hemolymph was up-regulated and reached the maximum level at 8 and 16 h, respectively, and then progressively dropped back to the original level. The results indicated that AISPI could play an important role in injury healing and immune response in mollusks as it could be induced by injury and bacterial challenge. (c) 2005 Elsevier Ltd. All rights reserved.

Molecular characterization and expression profiles in response to bacterial infection of Chinese soft-shelled turtle interleukin-8 (IL-8), the first reptilian chemokine gene

In this study, an IL-8 homologue has been cloned and identified from a reptile, Chinese soft-shelled turtle for the first time. The full-length cDNA of turtle IL-8 was 1188 bp and contained a 312 bp open reading frame (ORF) coding for a protein of 104 amino acids. The chemokine CXC domain, which contained Glu-Leu-Arg (ELR) motif and four cysteine residues, was well conserved in turtle IL-8. The 4924 bp genomic DNA of turtle IL-8 contained four exons and three introns. Phylogenetic analysis showed that the amino acid sequence of turtle IL-8 clustered together with birds. RT-PCR analysis showed that turtle IL-8 mRNA was constitutively expressed liver, spleen, kidney, heart, blood and intestine tissues of control turtles. Real-time quantitative PCR analysis further indicated that the turtle IL-8 mRNA expression was apparent in various tissues at 8 h and up-regulated significantly during 8 h-7 d after Aeromonas hydrophila infection. The present studies will help us to understand the evolution of IL-8 molecule and the inflammatory response mechanism in reptiles. (C) 2009 Elsevier Ltd. All rights reserved.

Gene expression profiles in liver of zebrafish treated with microcystin-LR

Microcystin-LR (MC-LR) is the most frequently studied cyclic heptatoxin produced by cyanobacteria, which has tremendous negative impacts on fish, while its molecular mechanism behind remained unclear at present. Here, Affymetrix Zebrafish GeneChip was used to identify alterations in gene expression of zebrafish (Danio rerio) after MC-LR exposure. Among the 14,900 transcripts in the microarray, 273 genes were differentially expressed, in which 243 genes were elevated and 30 were decreased. According to GOstat analysis, MC-LR mainly influenced the cell cycle and mitogen-activated protein kinases (MAPK) signaling pathways. In addition, many immune-related genes were also influenced. These data suggest that MC-LR could promote tumorigenesis and cause immunotoxicity in fish. (C) 2008 Elsevier B.V. All rights reserved.

Molecular cloning, promoter analysis and induced expression of the complement component C9 gene in the grass carp Ctenopharyngodon idella

Complement-mediated killing of pathogens through lytic pathway is an important effector mechanism of innate immune response. C9 is the ninth member of complement components, creating the membrane attack complex (MAC). In the present study, a putative cDNA sequence encoding the 650 amino acids of C9 and its genomic organization were identified in grass carp Ctenopharyngodon idella. The deduced amino acid sequence of grass carp C9 (gcC9) showed 48% and 38.5% identity to Japanese flounder and human C9, respectively. Domain search revealed that gcC9 contains a LDL receptor domain, an EGF precursor domain, a MACPF domain and two TSP domain located in the N-terminal and C-terminal, respectively. Phylogenetic analysis demonstrated that gcC9 is clustered in a same clade with Japanese flounder, pufferfish and rainbow trout C9. The gcC9 gene consists of 11 exons with 10 introns, spacing over approximately 7 kb of genomic sequence. Analysis of gcC9 promoter region revealed the presence of a TATA box and some putative transcription factor such as C/EBP, HSF, NF-AT, CHOP-C, HNF-3B, GATA-2, IK-2, EVI- 1, AP-1, CP2 and OCT-1 binding sites. The first intron region contains C/EBPb, HFH-1 and Oct-1 binding sites. RT-PCR and Western blotting analysis demonstrated that the mRNA and protein of gcC9 gene have similar expression patterns, being constitutively expressed in all organs examined of healthy fish, with the highest level in hepatopancreas. By real-time quantitative RT-PCR analysis, gcC9 transcripts were significantly up-regulated in head kidney, spleen, hepatopancreas and down-regulated in intestine from inactivated fish bacterial pathogen Flavobacterium columnare-stimulated fish, demonstrating the role of C9 in immune response. (c) 2007 Elsevier B.V. All rights reserved.