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国家高技术研究发展计划(2012AA10A402)

作品数:22 被引量:51H指数:4
相关作者:李军张志峰尤锋吴志昊郭华荣更多>>
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发文基金:国家高技术研究发展计划国家自然科学基金山东省自然科学基金更多>>
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栉孔扇贝心脏细胞的体外培养被引量:1
2018年
体外培养细胞对研究生物的资源保护、功能基因及病害发生机理与防治等均具有重要意义。然而,目前海洋双壳贝类中可以长期存活的组织细胞是有限的。本研究采用植块法启动栉孔扇贝(Chlamys farreri)心脏细胞的原代培养,通过优化培养基的方法,建立了可使其长期存活的原代培养体系。根据组织块迁出细胞的数量和细胞存活时间,确定L-15是3个培养基(L-15,M199,L-15+M199)中最适宜栉孔扇贝心脏细胞的基础培养基。通过三因子三水平正交实验,得出栉孔扇贝心脏细胞适宜的添加物组合:L-15基础培养基中添加5%胎牛血清、50 mmol/L牛磺酸和6 mmol/L Ca^(2+)。原代培养产物中以心肌细胞为主要的细胞类型,其中部分心肌细胞在原代培养14 d内可进行节奏性搏动;部分细胞可局部形成心肌束和肌管样结构;心肌细胞在体外存活时间达2个月。本研究将为栉孔扇贝基础生物学和功能基因的研究提供细胞平台。
季爱昌王华晏萌张志峰
关键词:栉孔扇贝心脏细胞体外培养正交实验
养殖牙鲆性类固醇激素的周年变化被引量:1
2013年
通过放射性免疫方法测定和比较了养殖牙鲆(Paralichthys olivaceus)成鱼血清中两种重要的性类固醇激素—睾酮(T)和雌二醇(E2)的周年变化规律。在雄性个体中,从4月份开始,随着水温的升高及繁殖期的开始,T和E2含量显著上升并在6月份达到最高峰;之后,T和E2含量逐渐下降并在第二年3月左右降低至最低水平。雌性牙鲆个体的E2含量于3月份开始慢慢增高,于6月份达到最高峰;随后,E2的含量逐渐降低并在1月份达到最小值;而雌性个体中的T的含量在2月和6月份出现两次高峰。结果表明,T和E2对牙鲆的性腺发育具有十分重要的作用,在雌雄个体中T和E2均呈现出周期性的变化。
孙鹏吴志昊尤锋李军
关键词:牙鲆性腺发育睾酮
Identification of Cytochrome P450 (CYP) Genes in Zhikong Scallop (Chlamys farreri)
2013年
Cytochrome P450 (CYP) superfamily is one of the membership largest and function most diverse protein superfamily recogniozed among living beings. Members of this superfamily were further assigned to different families and subfamilies based on their amino acid similarities. According to their phylogenetic relationships, the CYP genes which likely diverged from common ancestor gene and may share common functions were grouped into one clan. Widely distributing scallops are a group of the most conspicuous bivalve; however the studies on their CYP is acarce. In this study, we searched the genome and expressed sequence tags of Zhikong scallop (Chlamys farreri) for CYP genes. In total, 88 non-redundant CYP were identified, which were homed in 13 CYPs gene families. Phylogenetic analysis divided these genes into 4 CYP clans. As in deuterostomes, Clan 2 was the largest, which contained 33 genes belonging to CYP1, CYP2, CYP17 and CYP356 families. Clan 3 contgained 19 genes belonging to CYP3, CYP5 and CYP30 families. Clan 4 contained 23 genes, all belonging to CYP4 family. The mitochondrial CYP clan contained 9 genes belonging to CYP10 and CYP24 families. In comparison, protostomes (C. farreri, D. pluex, D. melanogaster) contained more CYP genes than deuterostomes (S. purpuratus and vertebrates) in Clan 2 but less genes in Clan 3 and Clan 4. Our findings will aid to deciphering CYP function and evolution in scallops and bivalves.
GUO HuihuiBAO ZhenminDU HuixiaZHANG LinglingWANG ShiSUN LuyangMOU XiaoyuHU Xiaoli
关键词:P基因CYP表达序列标签后口动物
piwi 1 is essential for embryogenesis and gametogenesis in scallop Chlamys farreri
Piwi,an animal-specific subclass of the Argonaute family have been known to involve in germline specification ...
Xiaoshi MaYunhan WangAichang JiZhifeng Zhang
关键词:EMBRYOGENESISGAMETOGENESIS
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条纹锯鮨精液超低温冷冻保存研究被引量:4
2014年
为建立条纹锯鮨精液超低温冷冻保存方法,实验采用计算机辅助精子分析系统(CASA)分析了采用6种抗冻保护剂(GLY[甘油]、DMSO[二甲基亚砜]、PG[丙二醇]、EG[乙二醇]、METH[甲醇]、DMA[二甲基乙酰胺])在4种浓度(5%、10%、15%、20%,v/v)下对条纹锯鮨精液的冷冻保存效果。结果发现,以HBSS为稀释液,采用程序降温仪分步降温冷冻保存条纹锯鮨精液,37℃水浴解冻后的精子中,15%PG作为抗冻保护剂的精子运动率最高,达到(93.1±0.9)%,与鲜精差异不显著(P>0.05),15%PG作为抗冻保护剂的精子水浴解冻后精子的运动速度最高,平均直线速度、平均曲线速度、平均路径速度分别达到了(88.3±0.3)μm/s、(76.2±0.5)μm/s、(86.7±0.7)μm/s,与鲜精差异不显著(P>0.05)。在不同种类及不同浓度抗冻保护剂保护下,15%PG作为抗冻保护剂的精子解冻后1 min内运动率变化与鲜精差异不显著(P>0.05)。研究表明,15%PG为条纹锯鮨最佳抗冻保护剂,可用于条纹锯鮨精液的超低温冷冻保存。
韩龙江刘清华官曙光纪利芹温海深李军
关键词:精子超低温保存
长牡蛎精子超低温冷冻后超微结构损伤研究被引量:9
2017年
采用程序降温仪分步降温冷冻保存长牡蛎(Crassostrea gigas)精液,并用扫描电镜、透射电镜研究了精子的超微结构损伤。超低温冷冻保存后长牡蛎精子的运动率、受精率及孵化率与鲜精无显著差异。鲜精中84.5%的精子形态结构正常,冻精中73%的精子形态结构正常。形态结构正常的精子表现为顶体、质膜、线粒体与鞭毛结构完整、染色质形状规则,顶体、线粒体及中心粒结构正常,鞭毛形态完整、微管结构清晰;形态结构异常的精子表现为顶体脱落、解体,精子头部质膜膨胀、破裂、染色质肿胀、破裂、解体,线粒体移位、脱落、膨胀,嵴退化或消失,鞭毛弯折、断裂,微管解聚。结果显示,以10%DMSO为抗冻保护剂,HBSS溶液为稀释液,1鲶4的稀释比例,添加海藻糖,采用分步降温法冷冻保存,对长牡蛎精子具有较好的抗冻保护作用,合适的冻存方法可以有效的保护太平洋牡蛎精子冷冻过程中结构损伤。研究有助于长牡蛎种质资源的收集保存及应用。
韩龙江刘清华许飞温海深李军
关键词:长牡蛎精子透射电镜超微结构
Development of 101 Novel EST-Derived Single Nucleotide Polymorphism Markers for Zhikong Scallop (Chlamys farreri)被引量:2
2013年
Zhikong scallop(Chlamys farreri) is an important maricultured species in China.Many researches on this species,such as population genetics and QTL fine-mapping,need a large number of molecular markers.In this study,based on the expressed sequence tags(EST),a total of 300 putative single nucleotide polymorphisms(SNPs) were selected and validated using high resolution melting(HRM) technology with unlabeled probe.Of them,101(33.7%) were found to be polymorphic in 48 individuals from 4 populations.Further evaluation with 48 individuals from Qingdao population showed that all the polymorphic loci had two alleles with the minor allele frequency ranged from 0.046 to 0.500.The observed and expected heterozygosities ranged from 0.000 to 0.925 and from 0.089 to 0.505,respectively.Fifteen loci deviated significantly from Hardy-Weinberg equilibrium and significant linkage disequilibrate was detected in one pair of markers.BLASTx gave significant hits for 72 of the 101 polymorphic SNP-containing ESTs.Thirty four polymorphic SNP loci were predicted to be non-synonymous substitutions as they caused either the change of codons(33 SNPs) or pretermination of translation(1 SNP).The markers developed can be used for the population studies and genetic improvement on Zhikong scallop.
LI JiqinBAO ZhenminLI LingWANG XiaojianWANG ShiHU Xiaoli
关键词:栉孔扇贝EST序列海水养殖品种等位基因
Association of myostatin Variants with Growth Traits of Zhikong Scallop(Chlamys farreri)被引量:2
2016年
Scallop is a popular sea food and an important aquaculture shellfish.Identification of genes and genetic variants relating to scallop growth could benefit high-yielding scallop breeding.Myostatin(MSTN) is a conservative regulator of muscle growth,and has become one of the most important target genes for genetic improvement of the production of farmed animals.In this study,four single nucleotide polymorphisms(SNPs) were identified in the 5' flanking region of MSTN gene(Cf MSTN) in Zhikong scallop(Chlamys farreri).The association of these SNPs with scallop growth traits,including shell length,shell height,body weight and striated muscle weight was analyzed.The SNP g-1162G>T was found to associate with shell length,shell height,and striated muscle weight.The TT type scallops showed significantly higher trait values than those of GT type,and the GG type individuals exhibited median values.On the contrary,significantly more Cf MSTN transcripts were detected in the striated muscle of GT type scallops than in those of TT and GG type ones.Our results suggested that Cf MSTN might regulate the scallop muscle growth negatively,and SNP g-1162G>T can be used as a candidate marker for the selective breeding of high-yielding scallop.
FU QiangGUO HuihuiFENG LiyingLI XueZHANG LinglingWANG ShiHU XiaoliBAO Zhenmin
关键词:肌肉生长抑制素性状变异MSTN
c-Myc基因在栉孔扇贝年周期发育性腺中的表达特征被引量:1
2019年
c-MYC在高等脊椎动物的细胞增殖、细胞分化和癌症发生中具有重要的调控作用。本研究克隆了栉孔扇贝cMyc基因的全长cDNA序列,分析了其在性腺年周期发育和配子发生过程中的表达特征。栉孔扇贝c-Myc cDNA全长2 943bp,可编码397个氨基酸,推导的氨基酸序列中除了包含MYC家族的保守结构域外,还具有c-MYC蛋白所特有的Myc-N转录激活区。qRT-PCR结果显示,栉孔扇贝卵巢中的c-Myc mRNA水平随着卵巢的发育和成熟显著升高;精巢中的表达水平在成熟期之前(休止期至生长期)逐渐升高,但在成熟期精巢中的表达水平显著低于增殖期和生长期。原位杂交和免疫组织化学结果显示,c-Myc mRNA及c-MYC蛋白的阳性信号均可在栉孔扇贝精巢的精原细胞和精母细胞以及卵巢的所有生殖细胞中检测到,推测其可能与栉孔扇贝的性腺发育和配子发生过程有关。
杨丹丹邓春杨乾坤秦贞奎张志峰
关键词:栉孔扇贝C-MYC配子发生性腺发育
对虾microRNA的最新研究进展
2018年
miRNA(microRNA)是一类广泛存在的长度约为18~25nt(nucleotides)的单链非编码小RNA。它在转录后水平通过降解靶基因的mRNA或者抑制靶基因mRNA的翻译来调控靶基因的表达,在生物的生长、发育、代谢和免疫等多种生命活动中发挥重要作用。对虾miRNA的研究起步较晚,但近6年来取得了突飞猛进的进展。本文综述了对虾miRNA的发现与鉴定、作用机制及其生物学功能方面的研究概况,并对对虾miRNA的研究前景进行了展望。
郭华荣郭华荣左建伟陈月如
关键词:对虾MIRNA靶基因
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