皱纹盘鲍对河流弧菌-Ⅱ苗免疫的研究

于1993-1995年，用免疫法防治皱纹盘鲍脓疱病。河流弧菌-Ⅱ分离自大连水产养殖公司、太平洋海珍品养殖公司的患脓疱病的皱纹盘鲍;健康皱纹盘鲍，由大连水产养殖公司石庙苗种厂提供。鲍血淋巴细胞的体外吞噬实验:取健康皱纹盘鲍的血淋巴与肝素和菌液按一定比例混合后，置湿盒内37℃培养30min，取一滴涂片，吕氏液染色后观察。用该病原菌制做菌苗，大量扩增病原菌，用0.1％-1.0％的甲醛洗下菌苔，处理24-120h，用生理盐水反复清洗，除掉甲醛;再用生理盐水稀释到所需浓度。通过反复传代降低毒性，制成活菌苗。用两种菌苗在稚鲍、一龄鲍（2-3cm）和成鲍（6-8cm）上实验。结果表明，鲍的血淋巴细胞约有三种，三种细胞都有吞噬细菌的能力，有的细胞象变形虫样变形，在细胞膜上吸附着很多细菌，有的细菌已被吞到细胞内;该病原菌可同鲍的血清发生凝集反应;菌苗的效果明显，可提高成活率约50％以上。由上述结果可见，皱纹盘鲍的血淋巴细胞具有吞噬细菌的能力，在菌苗的作用下，血淋巴细胞增殖并活跃地吞噬异源致病菌，同时鲍血液中的免疫因子也被激活，参与免疫的过程，使皱纹盘鲍成活率明显提高。     

几种动物精卵识别机制

动物受精是指两性配子细胞相结合生成合子的过程,亲代通过受精将遗传物质传给子代,它是衔接亲代与子代的桥梁,保持了物种的延续.精卵识别是受精的起始步骤,是指配子细胞通过各自表面的受精蛋白--糖蛋白类物质的特异性相互作用识别并结合,完成受精过程.本文着重介绍目前研究比较清楚的小鼠、猪、海胆和鲍等的精卵识别机制,比较了哺乳动物和无脊椎动物之间的联系和差别,进一步说明不同动物通过各自性细胞表面的糖蛋白完成精卵识别过程,并由相应糖蛋白的不同组成和结构决定了该识别过程的特异性.     

Construction of a cDNA library for sea cucumber <Emphasis Type="Italic">Acaudina leucoprocta</Emphasis> and differential expression of ferritin peptide

Acaudina leucoprocta is an edible sea cucumber of economic interest that is widely distributed in China. Little information is available concerning the molecular genetics of this species although such knowledge would contribute to a better understanding of the optimal conditions for its aquaculture and its mechanisms of defense against disease. Therefore, we constructed a cDNA library and, based on bioinformatics analysis of the sequences, the functions of 75% of the cDNAs were identified, including those involved in cell structure, energy metabolism, mitochondrial function, and signal transduction pathways. Approximately 25% of genes in the library were unmatched. The gene for A. leucoprocta ferritin was also cloned. The predicted amino-acid sequence of ferritin displayed significant homology with other sea-cucumber counterparts but indicated that it was a new member of the ferritin family. Semiquantitative real-time RT-PCR indicated the highest levels of ferritin mRNA expression in the intestine. A polyclonal antibody of ferritin was also produced. These data provide a set of molecular tools essential for further studies of the functions of ferritin protein in A. leucoprocta.     

虾夷马粪海胆黑嘴病的初步研究

对虾夷马粪海胆“黑嘴病”进行了初步的研究。经分离、纯化后鉴定黑嘴病的病原菌为坚强芽孢杆菌 (BacillusfirmusBredemannetWarner)。同时报道了该菌为革兰氏阳性杆菌 ,大小为(0.6～0.7)μm×(0.8～1.0)μm。可在1%～7%NaCl中生长 ,最适盐度2～3 ;最适pH为7～8;生长温度为8～26℃ ,最适生长温度为15℃。该菌对磺胺甲基异恶唑、复方新诺明和头孢噻肟等抗生素敏感 ,可用之治疗     

糖蛋白及其在动物精卵识别中的作用

糖蛋白是指由比较短 ,往往带分支的寡糖与多肽链共价连接而成的一类结合蛋白质。其中 ,糖部分所占比例较小。糖蛋白作为生物体内重要的生物大分子 ,其种类繁多 ,并且以各种形式广泛存在于细胞膜、细胞间质、血浆以及粘液中 ,具有开关、调谐、作为激素、胞内转运、保护、促进物质吸收、参与血液凝固、参与细胞识别等多方面的重要生理活性。1糖蛋白的组成糖蛋白分子是由多肽链和糖两部分组成的 ,糖和多肽链之间通过糖苷键相连接。其中的蛋白质是生理功能的主要担负者 ,糖链起到维持蛋白质结构稳定 ,抵抗蛋白酶水解 ,防止抗体识别及参与多肽链…     

泥蚶(Tegillarca granosa)精氨酸激酶的基因克隆与表达

利用PCR扩增技术,从泥蚶的cDNA文库中克隆出了精氨酸激酶(arginine kinase)基因,并进行了原核表达.精氨酸激酶是调节能量代谢的重要酶类,在无脊椎动物体内能量平衡过程中起重要作用.结果表明,泥蚶的精氨酸激酶全长1601bp,包括5'非翻译区(5'-UTR)序列332bp,3'非翻译区(3'-UTR)序列...     

可口革囊星虫(Phascoloma esculenta)铁结合蛋白基因的研究

根据已报道的铁结合蛋白基因的保守序列设计引物,用cDNA末端快速扩增(RACE)法,成功地从可口革囊星虫体液中获得铁结合蛋白基因的全长序列.结果表明,该基因cDNA全长1017bp,5'-端非编码区为15lbp,3'一非编码区为341bp,开放阅读框长度为525bp(括一个终止密码子),可编码175个氨基酸(GenBank:EU091352).该序列与加洲海兔、皱纹盘鲍、刺参、微小牛蜱、叉尾鲴、非洲爪蟾、小家鼠、人等的铁结合蛋白基因有67%-75%的同源性,而相应的氨基酸同源性为59%-76%.氨基酸相似性为74%*-89%%.分析结果表明,铁结合蛋白基因在动物进化中是高度保守的.     

Isozyme expression of Chinese and Japanese populations of Chlamys farreri and their reciprocal hybrids

Chinese and Japanese population of Chlamys farreri and their reciprocal hybrids were surveyed in isozyme variability at 13 loci by polyacrytamide gel electrophoresis (PAGE). Isozyme banding patterns indicated these hybrids were diploid. Loci that were observed as being monomorphic in inbred populations of C. farreri were also found to be monomorphic in filial progeny; loci that observed to be polymorphic in parental type generations were also polymorphic in hybrid generations. Differences existed among allelic frequency of the four types of cross. Within the reciprocal hybrids the expression of malic enzyme (ME) isozyme was sufficient to distinguishing individual hybrids because of the band, Rf=0.38. However, there were no noticeable variations among all the samples to differentiate one from another. Inbreeding was likely to be the main problem in aquaculture. The introduction of new broodstock can improve the genetic diversity. Hybrid vigor has manifested to a certain extent in the present study.     

可口革囊星虫(Phascolosoma esculenta)酵母双杂交文库和铁结合蛋白真核表达载体的构建

构建了可口革囊星虫(Phascolosoma esculenta)的酵母双杂文库, 总克隆数为6.09×106CFU, 重组率100%, 平均插入片段长度>1kb。随机挑取200个克隆子测序, 结果1个空载, 67 个无法对应的基因类型, 已知的132个基因中线粒体蛋白占29.54%, 蚯蚓血红蛋白15.9%, 核糖体蛋白10.6%, 铁结合蛋白8.3%, 肌动蛋白6.1%, 其它基因29.56%。这些基因分别为: 5-氨基乙酰丙酸合成酶、类博莱霉素水解酶、纤溶蛋白、谷氨酰胺合成酶、热休克蛋白、非特性类碱性磷酸酶、脂肪酶、金属蛋白、线粒体、硫氧还蛋白、蛋白磷酸酶、肌钙蛋白C、泛素。根据已获得的可口革囊星虫铁结合蛋白基因, 以EcoRⅠ和KpnⅠ为双酶切位点设计引物, 构建了可口革囊星虫铁结合蛋白真核表达载体pPink-HC-fer, 经PCR 及测序检验, 序列完全正确, 然后电转化至酵母细胞中诱导表达, 经SDS-PAGE检验其融合蛋白分子量大小约为23kDa。