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Identification and Expression of the Conotoxin Homologous Genes in the Giant Triton Snail (Charonia tritonis)
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作者 JIA Huixia ZHANG Gege +4 位作者 ZHANG Chenglong ZHANG Hua YAO Gaoyou HE Maoxian LIU Wenguang 《Journal of Ocean University of China》 SCIE CAS CSCD 2023年第1期213-220,共8页
Conotoxin(CTX)is a small peptide toxin,and plays crucial role in anesthetizes prey,preys and defends competitors.Charonia tritonis is the natural predator of the crown of thorns starfish(CoTS)and the protector of cora... Conotoxin(CTX)is a small peptide toxin,and plays crucial role in anesthetizes prey,preys and defends competitors.Charonia tritonis is the natural predator of the crown of thorns starfish(CoTS)and the protector of coral reefs.It plays an important role in the coral reef ecosystem.However,the types of toxins produced by tritons and the molecular mechanisms of toxin secretion of C.tritonis are unknown.In the present study,the four conotoxin homologous genes(CTXs)from C.tritonis were identified.Species and conotoxin superfamily phylogenetic tree indicated that CTX-1(CL2216.Contig2)and CTX-4(Unigene 58438_All)belong to the C superfamily,CTX-2(Unigene 66414_All)belong to the V superfamily,and CTX-3(Unigene 21408_All)belong to the B1 superfamily.CTXs were highly expressed in salivary gland,liver and digestive gland.The predation control experiment revealed that the expressions of CTXs were significantly different in salivary gland,liver and digestive gland.The results indicated that CTXs may participate in the process of C.tritonis predating CoTs,and provided a scientific basis for further studying the toxins secretion mechanism of C.tritonis. 展开更多
关键词 charonia tritonis TOXIN tissue distribution predation experiment
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Cryopreservation of Veliger Larvae of Trumpet Shell, Charonia sauliae: an Essential Preparation to Artificial Propagation 被引量:1
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作者 KANG Kyoung Ho ZHANG Zhifeng +1 位作者 BAO Zhenmin SHAO Mingyu 《Journal of Ocean University of China》 SCIE CAS 2009年第3期265-269,共5页
Trumpet shell, Charonia sauliae, is an endangered and valuable species, but its artificial propagation protocol has not been successfully established. To estimate the possibility of cryopreservation for larvae of C. s... Trumpet shell, Charonia sauliae, is an endangered and valuable species, but its artificial propagation protocol has not been successfully established. To estimate the possibility of cryopreservation for larvae of C. sauliae, which is a potential preparation for its artificial reproduction and further research, in this study a protocoi for the cryopreservation of veliger larvae of trumpet shell was optimized. Through a two-step cryopreservation procedure, four kinds of cryoprotectants (ethylene glycol, 1, 2-propanediol, dimethyl sulfoxide and glycerol) were employed at three concentrations (1.0, 1.5 and 2.0molL^-1) respectively and survival rates of larvae were determined after a storage of lh. The larvae frozen with these four cryoprotectants after 1 h storage were cultured, and then survival rates were determined at 24, 72 and 120 h after thawing. Dimethyl sulfoxide at a concentration of 1.5 molL^-1 showed the best protective effect in all experiments (p〈0.05). And survival rates of larvae frozen with dimethyl sulfoxide were determined after 1, 7 and 15 d of storage. The survival rates of larvae frozen with 1.5 molL^-1 dimethyl sulfoxide after 1 h, 1 d, 7 d and 15 d of storage were 80.77% ±7.51%, 80.34%±11.28%, 83.10%±9.14% and 77.23%±6.22% respectively. No significant differences in survival rates of larvae frozen with dimethyl sulfoxide were observed after various storage periods (p〉0.05). 展开更多
关键词 trumpet shell charonia sauliae veliger larva CRYOPRESERVATION
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Feeding Broodstocks Different Starfish Diets Affect Growth and Survival of Larvae of Trumpet Shell(Charonia lampas sauliae Reeve 1844)
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作者 KANG Kyoungho KIM Malhee +2 位作者 HONG Hyeran CHA Guyong SUI Zhenghong 《Journal of Ocean University of China》 SCIE CAS 2016年第5期861-865,共5页
Trumpet shell(Charonia lampas sauliae)(Mollusca, Heterogastropoda, Cymatidae) has extensive economic value. Studies on the artificial larval development of C. lampas sauliae for aquaculture utilization have become esp... Trumpet shell(Charonia lampas sauliae)(Mollusca, Heterogastropoda, Cymatidae) has extensive economic value. Studies on the artificial larval development of C. lampas sauliae for aquaculture utilization have become especially important due to the finite natural resources. In the present study, the growth and survival rate of the larvae of C. lampas sauliae broodstocks fed three types of starfish diets, Asterina pectinifera Müller & Troschel 1842, A. amurensis Lütken 1871 and their mixture were compared. The larval size increased gradually between day 10 and day 20 after hatching at 15℃ and 20℃. No difference was found in body size and specific growth rate(SGR)(two-way ANOVA; P > 0.05). However, during transition from trochophore to veliger stage 20 days after hatching, significant increases in larval survival and growth rates were observed. The maximum survival rate was observed on day 10. The mean survival rate was 0.463, 0.730 and 0.515 at 15℃, and 0.369, 0.713 and 0.444 at 20 when ℃ A. pectinifera, A. amurensis and their mixture were fed, respectively. The SGR and survival rate of the larvae were definitely influenced by the diets(P < 0.05), and the effect of A. amurensis alone was higher than that of A. pectinifera alone and their mixture. 展开更多
关键词 larvae Mollusca aquaculture definitely alone length capsules seawater cilia filtered
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基于RNA-Seq高通量测序技术的法螺差异表达基因分析
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作者 刘文广 张格格 +1 位作者 岑希彤 何毛贤 《热带海洋学报》 CAS CSCD 北大核心 2023年第4期146-154,共9页
法螺是珊瑚礁“杀手”长棘海星为数不多的自然捕食者之一,是珊瑚礁生态系统的重要保卫者。但是,由于海洋环境变化、人类的过度捕捞、珊瑚礁栖息地被破坏等因素,法螺的自然资源遭受严重破坏,已长期处于濒临灭绝的状态。尽快开展法螺基础... 法螺是珊瑚礁“杀手”长棘海星为数不多的自然捕食者之一,是珊瑚礁生态系统的重要保卫者。但是,由于海洋环境变化、人类的过度捕捞、珊瑚礁栖息地被破坏等因素,法螺的自然资源遭受严重破坏,已长期处于濒临灭绝的状态。尽快开展法螺基础研究迫在眉睫,但是该物种的基因组资源有限,阻碍了如生物矿化等分子机制的深入研究。本研究借助于Illumina HiSeq^(TM)2000测序平台完成了法螺两种组织(肌肉与外套膜)的高通量测序工作。测序所得原始序列经筛选、组装之后比对到NR、NT、Swissprot、KEGG、GO和InterPro数据库中获得注释结果,并进行差异基因聚类分析。结果表明,测序共获得109722个Unigene,其中有46239个Unigene得到功能注释,共获得7994个差异表达基因,其中在法螺外套膜组织中有3196个上调,4798个下调。对差异表达基因(differentially expressed genes,DEG)的GO注释和KEGG富集分析显示,高表达的DEG显著富集在典型的生物矿化相关通路中,如糖胺聚糖生物合成硫酸软骨素通路(glycosaminoglycan biosynthesis chondroitin sulfase dermatan sulfate)、磺基转移酶活性(sulfotransferase activity)、钙离子结合(calcium ion binding)等。通过实时定量荧光聚合酶链反应(RT-qPCR)对8个与矿化相关的DEG基因的表达情况进行验证表明,其表达趋势与转录组测序结果一致。本研究中所获得丰富的转录本和全面的转录组信息,增加了法螺基因信息的同时,还为法螺矿化机制的解析提供了一定的数据支持。 展开更多
关键词 法螺 转录组 差异表达基因 生物矿化
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法螺G型溶菌酶重组蛋白的原核表达及抑菌活性探究
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作者 贾慧霞 张华 +2 位作者 张成龙 何毛贤 刘文广 《海洋科学》 CAS CSCD 北大核心 2022年第10期32-42,共11页
G型溶菌酶(G-type lysozyme,G.lys)是一种富含半胱氨酸的天然免疫因子,在无脊椎动物的免疫防御过程中起重要作用。本研究旨在通过揭示法螺(Charonia tritonis)G型溶菌酶(CtG.lys)重组蛋白的抑菌功能,为进一步认识法螺的分子免疫机理及... G型溶菌酶(G-type lysozyme,G.lys)是一种富含半胱氨酸的天然免疫因子,在无脊椎动物的免疫防御过程中起重要作用。本研究旨在通过揭示法螺(Charonia tritonis)G型溶菌酶(CtG.lys)重组蛋白的抑菌功能,为进一步认识法螺的分子免疫机理及病害防治提供新思路。首先对CtG.lys序列进行了分析,利用实时荧光定量PCR检测CtG.lys的组织分布,然后以pGEX-4T-1为表达载体,构建重组质粒pGEX-4T-1-CtG.lys,进行重组蛋白表达,并检测了重组蛋白的抑菌活性。结果显示:1)CtG.lys cDNA的5′非翻译区(UTR)为66 bp,3′UTR为186 bp,开放阅读框为789 bp,编码262个氨基酸,有1个可溶性转糖苷酶(SLT)结构域(72~255 aa),包含6个半胱氨酸残基和3个酶活性催化位点(Glu72、Asp85、Asp96);2)多序列比对及系统发育分析表明,CtG.lys与亲缘关系较近的福寿螺G型溶菌酶氨基酸的同源性最高;3)CtG.lys在所有被检测组织中均表达,其中在肝脏、外套膜表达量较高;4)16℃,0.5 mmol/L IPTG诱导表达10 h后,重组蛋白在上清液和包涵体均表达分子量约为51.91 kDa,纯化后得到浓度为1.2 mg/mL的重组蛋白;5)抑菌实验表明,重组蛋白pGEX-4T-1-CtG.lys具有抗金黄色葡萄球菌Staphyloccocus aureus、藤黄微球菌Micrococcus luteus、施罗氏弧菌Vibrio shilonii的活性,对革兰氏阳性细菌的抑菌作用大于革兰氏阴性细菌。研究结果为进一步解析法螺免疫防御的分子机理提供了参考数据和科学依据。 展开更多
关键词 法螺 G型溶菌酶 原核表达 抑菌活性
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法螺线粒体全基因组密码子偏好性分析 被引量:2
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作者 任童童 喻达辉 +5 位作者 翟子钦 谭传港 何积翠 尚东维 陈移波 白丽蓉 《南方农业学报》 CAS CSCD 北大核心 2022年第5期1379-1387,共9页
【目的】分析法螺(Charonia tritonis)线粒体基因组密码子偏好性,探究基因突变和自然选择对密码子偏好性的影响,为法螺属动物类群的系统发育树构建及种质资源保护与遗传改良提供参考依据。【方法】以法螺线粒体全基因组序列为材料,选择... 【目的】分析法螺(Charonia tritonis)线粒体基因组密码子偏好性,探究基因突变和自然选择对密码子偏好性的影响,为法螺属动物类群的系统发育树构建及种质资源保护与遗传改良提供参考依据。【方法】以法螺线粒体全基因组序列为材料,选择以ATG为起始密码子的非重复且长度大于300 bp的11个CDS(Coding DNA sequence)序列,采用CodonW 1.4.2进行密码子偏好性分析,并根据相对使用度差值(△RSCU)>0.08且RSCU在高表达基因库中>1.00的标准筛选出最优密码子。【结果】法螺线粒体基因组密码子第1位GC含量(GC1)的波动范围在35.80%~53.10%,平均为43.54%;第2位GC含量(GC2)的波动范围在32.40%~43.40%,平均为36.11%;第3位GC含量(GC3)的波动范围在23.00%~30.60%,平均为27.79%,GC1和GC2明显高于GC3;同义密码子第3位的GC含量(GC3_(S))的波动范围在20.90%~29.60%,平均为26.23%,表明密码子第3位碱基以A/U(T)结尾为主;密码子适应指数(CAI)的波动范围为0.096~0.166,平均为0.137;密码子偏好性指数(CBI)的波动范围在-0.212~-0.086,平均为-0.133;最优密码子使用频率(FOP)的波动范围为0.246~0.339,平均为0.306;有效密码子数目ENC的波动范围在44.37~51.69,平均为47.84;总平均亲水性(GRAVY)的变化范围在0.4419~1.4727,平均为0.9501。中性绘图分析、ENC-plot绘图分析及对应性分析均表明,在法螺线粒体基因组密码子偏好性的形成过程中受基因突变和自然选择的双重作用,但以自然选择为主导。综合高表达优越密码子和高频密码子,最终确定出11个密码子为法螺线粒体基因组最优密码子,分别为UUU、CUA、GUA、UAU、CAA、AAA、UCA、CCA、ACA、GCC和GCA,其中有1个密码子以G/C结尾,其余10个密码子均以A/U结尾。【结论】法螺线粒体基因组密码子偏好以A/U(T)结尾,筛选确定的11个最优密码子主要以A/U结尾。法螺线粒体基因组密码子为弱偏好性密码子,自然选择在其偏好性的形成过程中起主导作用。 展开更多
关键词 法螺 线粒体基因组 密码子偏好性 最优密码子 自然选择
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