Purification and identification of a clotting protein from the hemolymph of Chinese shrimp (Fenneropenaeus chinensis)

The clotting protein (CP) plays important and diverse roles in crustaceans, such as coagulation and lipid transportation. A clotting protein was purified from the hemolymph of Chinese shrimp Fenneropenaeus chinensis (named as Fc-CP) with Q sepharose HP anion-exchange chromatography and phenyl sepharose HP hydrophobic interaction chromatography. Fc-CP was able to form stable clots in vitro in the presence of hemocyte lysate and Ca²⁺, suggesting that the clotting reaction is catalyzed by a Ca²⁺-dependent transglutaminase in shrimp hemocytes. The molecular mass of Fc-CP was 380 kDa under non-reducing conditions and 190 kDa under reducing conditions as was determined with SDS-PAGE. CP exists as disulfide-linked homodimers and oligomers. The N-terminal amino acid sequence of Fc-CP was identical to that of shrimps including Penaeus monodon, Farfantepenaeus paulensis and Litopenaeus vannamei; and similar to that of other decapods. The purified Fc-CP was digested with trypsin and verified on an ABI 4700 matrix-assisted laser desorption/ionization tandem time-of-flight (MALDI-TOF/TOF) mass spectrometry. Our results will aid to better understanding the coagulation mechanism of shrimp hemolymph.     

Construction of a muscle cDNA library of Chinese shrimp <Emphasis Type="Italic">Fenneropenaeus chinensis</Emphasis> and sequence analysis of the troponin I gene

A muscle cDNA library of Chinese shrimp (Fenneropenaeus chinensis) was constructed with the SMART™ cDNA Library Construction Kit. The titer of optimal primary library was 7.7×10⁵ pfu mL⁻¹ and that of the amplified library was 3.0×10⁹ pfu mL⁻¹. The percentages of the recombinant clones of primary and amplified libraries were over 98%. The insert sizes were longer than 400 bp with an average of 1000 bp. A positive clone containing a 794 bp insert was sequenced and identified encoding fast skeletal troponin I gene. This library provided a useful resource for the functional genomic research of F. chinensis.     

一种新的中国对虾弧菌病原菌──产气弧菌

于１９９３年８月，从山东省昌邑县下营镇患败血病的中国对虾血淋巴内分离到多株细菌，其中２株人工感染证实为病原菌；经５０多项形态，生理，生化特性测试鉴定为产气弧菌Ｖｉｂｉｏｇａｚｏｇｅｎｅｓ。其主要特性为：革兰氏阴性，弧状，极生单鞭毛，产生红色素，氧化酶呈阳性，精氨酸双水解酶，赖氨酸脱羧酶，鸟氨酸脱羧酶均为阴性，淀粉酶，明胶酶为阳性，硝酸盐还原阴性，利用木糖，水杨苷，山梨醇发酵葡萄糖产气，利用柠檬酸盐     

中国对虾卵母细胞发育的初步研究

于１９８９年１０月－１９９０年９月分别先后在昌邑县下营镇对虾育苗场和青岛黄岛盐业公司养殖场采集中国对虾，运用光学，电子显微镜及组化方法，研究中国对虾卵母细胞的形态发育及其内部结构分化。结果表明，中国对虾卵母细胞的发育可分为４个主要时相：Ⅰ．早期分化时相，Ⅱ滤泡初生期时相，Ⅲ卵黄大量生成时相，Ⅳ皮质棒生成时相。     

答《关于深海锰结核中“纳米级超微生物化石”的思考和建议》一文

本文详细讨论了东太平洋深海锰矿结核中新发现的中华微放线菌和太平洋螺球孢菌究竟是不是超微结构，首先，它们是笔者根据形态，结构和元素成分作为超微生物化石进行鉴定的，其次，透射电子显微镜中可以清楚地区分出完整的微生物结构，包括菌丝，孢子，孢囊和菌落等，它们之间的尺寸比例也是适当的，此外，透射电子显微像还揭示，深海锰质叠层石的纹层就是这些超微生物韵律性生长的结果，与滨海叠层中的纹层十分相似，笔者认为，已知微生物的最小尺寸不能作为否定新发现的超微生物化石的依据。本文还讨论了超微生物化石的术语问题。     

The Complete Sequence of Mitochondrial COⅡ Gene of Fenneropenaeus chinensis and Its Applicability as a Marker for Phylogenetic Analysis

The complete mitochondrial cytochrome oxidase subunit Ⅱ (COⅡ) gene of Penaeinae shrimp Fenneropenaeus chinensis was cloned and sequenced. The gene is 688 bp in length and codes for 229 amino acids. It shows 83.2%, 87.0% and 83.8% sequence similarity to Marsupenaeus Japonicus, Penaeus monodon and Farfantepenaeus notialis, respectively. The A+T content of the whole gene and that at the third position of codons are 64.7% and 78.2%, respectively. The phylogenetic relationship between F. chinensis and three other species representing genera Farfanatepenaeus, Marsupenaeus and Penaeus was analyzed. Results showed that the genetic distances among the four taxa ranged from 0.144 0 to 0.200 5, exceeding those estimated with COⅠ and partial 16S rRNA gene sequences among Marsupenaeus, Litopenaeus and Melicertus, and being therefore larger than the value among subgenera. It has been suggested that the COⅡ gene has a faster evolutionary rate than that of the COⅠ gene and partial 16S rRNA gene and could be used for phylogenetic analysis at genus or species level. The results of the present study indicated that Farfantepenaeus, Fenneropenaeus, Marsupenaeus and Penaeus are at a higher phylogenetic level than subgenus, which supports the opinion of the elevation of phylogenetic status of the four subgenera to genus level.     

中国对虾(Fenneropenaeus chinensis)视神经节神经内分泌细胞电生理特征

利用膜片钳技术对体外培养24—48h中国对虾的视神经节端髓A型和B型神经内分泌细胞进行电生理特征研究。电流钳的结果表明，A型细胞具有自发和诱发放电活动；B型细胞没有记录到明显放电活动。在电压钳模式下，中国对虾视神经节神经内分泌细胞表达TTX敏感Na^ 通道、高电压激活L型Ca^2 通道、TEA敏感晚钾通道、早钾通道。Ca^2 通道电流在-40— -30mV被激活，在-10—0mV时达到峰值；Ca电流受钳制电压的影响。一定电压范围内，钳制电压越负，Ca电流就越大。外向的钾通道电流均在-40mV左右被激活。晚钾通道电流对TEA敏感，但TEA不能完全阻断晚钾通道电流。     

Application of RAPD technology for identification in three different stocks of Penaeus chinensis

INTRODUCTIONPenaeuschinensishasbecomeoneoftheprimaryspeciesforaquacultureinChina .How ever ,viralandbacterialdiseaseproblemshaveplaguedtheshrimpaquacultureindustryserious lyinChinasince 1 993.Ithasbeenindicatedthatthereducedgrowthperformanceandthein creasedsusceptibilitytodiseasescouldbearesultoflowlevelsofgeneticvariablities (Carretal .,1 994 ) .InordertosupporttheaquacultureindustryoftheUnitedStates,theUSMarineShrimpFarmaringProgramConsortiumhasdevelopedvariousdisease free populati…     

利用WSBV的引物扩增中国对虾的杆状病毒DNA

利用台湾报道的ＷＳＢＶ的ＰＣＲ引物,成功地扩增出了１４００ｂｐ左右的中国对虾的杆状病毒的ＤＮＡ片段,与预计的产物长度吻合。该引物对经初步离心处理的病虾组织也能扩增出特异性ＤＮＡ条带,而对正常虾组织ＤＮＡ、昆虫杆状病毒（ＡｃＭＮＰＶ）ＤＮＡ进行扩增,均获得阴性结果。说明该引物的特异性较高,对中国对虾的杆状病毒的检测具有一定的实际应用价值。实验结果同时说明ＷＳＢＶ与中国对虾的杆状病毒有同源序列存在,具有一定的保守性;用该引物进行的ＰＣＲ扩增为中国对虾的杆状病毒与其他相关病毒的比较研究提供了一种技术手段。     

摄食对中国对虾能量代谢影响的初步研究

于 1996年 5～ 6月在水温 2 5℃时测定了两种规格中国对虾 ( Penaeus chinensis)摄食沙蚕和配合饲料后的耗养率和氨排泄率。小规格中国对虾摄食配饵的代谢率略高于摄食沙蚕 ,而大规格组中摄食沙蚕的代谢率大于摄食配饵组。小规格中国对虾摄食配饵和沙蚕的代谢能比未摄食组提高了 33.2 2 %和 2 7.4 0 %;大规格中国对虾摄食配饵和沙蚕的代谢能比未摄食组提高了 34 .78%和 4 0 .0 5%。中国对虾摄食后的氨排泄率明显增加 ,表明蛋白质在摄食代谢中具有重要作用。