磷酸泛酰巯基乙胺基转移酶基因过量表达对集胞藻PCC6803脂肪酸合成的影响

磷酸泛酰巯基乙胺基转移酶（phosphopantetheinyl transferases,PPTase）是细菌脂肪酸合成中的关键酶。本研究利用同源重组手段构建集胞藻PPTase基因（slr0495）过量表达重组质粒,在集胞藻PCC6803中进行表达研究,并在DNA和RNA水平进行验证,利用气相色谱-质谱联用仪（GC-MS）检测不同条件下突变藻株脂肪酸组分及含量。结果表明：在温度为30 ℃、光照强度为50 μmol ·m^-2 · s^-1培养条件下,过量表达slr0495基因突变藻株中C12:0、C16:0和C18:0的含量分别为1. 31 mg · g^-1、8.07 mg · g^-1和1.35 mg · g^-1,比野生型藻株分别提高了23.58%、25.31%和13.45%;在温度为20 ℃、50% NaNO₃浓度、光照强度为50 μmol · m^-2 · s^-1培养条件下,与野生型相比,突变藻株中C16:0和C18:0含量分别提高了44.71%和41.51%。以上结果表明,过表达slr0495基因增加了集胞藻PCC6803中长链饱和脂肪酸的含量,并且在低温（20 ℃）和缺氮（50% NaNO₃）双重胁迫培养条件下中长链饱和脂肪酸含量得到进一步提高。本研究为探究slr0495基因功能以及在逆境中基因产生的应激反应提供了理论依据,为微藻高产多不饱和脂肪酸代谢研究奠定了基础。     

Effects of salinity and temperature during incubation on hatching and development of lingcod Ophiodon elongatus Girard, embryos

The interactive effects of salinity and temperature on development and hatching success of lingcod, Ophiodon elongatus Girard, were studied by incubating eggs at four temperatures (6, 9, 12 and 15°C) and five salinities (15, 20, 25, 30 and 35 g L^?1). Hatch did not occur in any of the 15°C treatments. Degree days (°C days) to first hatch was not influenced by temperature or salinity, however, calendar days to first hatch differed significantly for temperature (P<0.0001, 61±1, 44±1 and 35±1 days for 6, 9 and 12°C respectively). Degree days to 50% (427.1±4.2) hatch was not significantly influenced by temperature but was by salinity (P=0.0324). Viable hatch (live with no deformities, 74.1±4.0%) was greatest at 9°C and 25 g L^?1 but not significantly different in the range of 20–30 g L^?1. Larval length (9.4±0.13 mm) was greatest at 9°C and 20–30 g L^?1. Temperature and salinity significantly influenced all categories of deformities with treatments at the upper (12°C and 35 g L^?1) and lower limits (6°C and 15 g L^?1) producing the greatest deformities. The optimal temperature and salinity for incubating Puget Sound lingcod eggs was found to be 9°C and 20–30 g L^?1.     

Host preference of adult Caligus elongatus Nordmann in the laboratory and its implications for Atlantic cod aquaculture

The sea louse Caligus elongatus utilizes a range of marine fish species as hosts, and it has been suggested that fish farms are infected with adult C. elongatus from wild fish hosts. We assessed experimentally whether dislodged lice return to their original host species. Adult lice from wild lumpfish and saithe were presented to lumpfish, sea trout, Atlantic cod, saithe and European plaice. All fish species became infected. Lumpfish and cod were strongly preferred, followed by saithe, trout and plaice. Lice from wild saithe showed a preference for saithe and lumpfish, followed by cod, sea trout and plaice. The mtCO1 gene was used to determine the genotypes of the lice. Lice from lumpfish were genotype 1. Lice from saithe were mainly genotype 2. The two genotypes of C. elongatus differed slightly in their host preferences: lice from saithe were less discriminating. The data support the hypothesis that adult C. elongatus may transfer between different host species. Cod is an attractive host for lice from both lumpfish and saithe, which suggests that C. elongatus may become a serious pest of farmed cod.     

Sea lice treatments, management practices and sea lice sampling methods on Atlantic salmon farms in the Bay of Fundy, New Brunswick, Canada

Proper monitoring of all stages of sea lice is imperative to ensure the strategic timing of treatments. Although sea lice receive close attention and management for production purposes, there are no regulations for the reporting of lice burdens on salmon farms in Atlantic Canada, nor are there officially standardized protocols for conducting sea lice counts in the field. The purpose of this study was to complete a survey of Atlantic salmon farms in the Bay of Fundy, NB, Canada. The survey addressed many aspects of sea lice monitoring and control, including methods for sea lice sampling at the site, types of treatments administered for control of sea lice and management practices and decisions regarding the control of sea lice. A total of 83 surveys were completed from July to December 2002. Twelve of the 60 respondents owned and/or operated more than one site and completed a survey for each of their sites at the time of the interview. The position of individuals surveyed included 48 site managers (representing 63 sites), three site owners (representing 11 sites), eight individuals who were both owner and manager (representing nine sites) and one site veterinarian (representing one site). Surveys were completed on 30 sites from the Limekiln Bay and surrounding areas, 23 sites from Grand Manan Island, 18 sites from Deer Island, six sites from Campobello Island and six sites from Passamaquoddy Bay. Fish farmers, in concert with their veterinarians, generally perform intensive monitoring and control practices. However, there is a continued reliance on emamectin benzoate (SLICE^®) for sea lice control on Atlantic salmon farms in the Bay of Fundy, which raises concerns regarding the potential for sea lice to develop resistance to the drug.     

Role of free neuromasts in larval feeding of willow shiner Gnathopogon elongatus caerulescens Teleostei, Cyprinidae

ABSTRACT:   It has been reported that the larvae of willow shiner Gnathopogon elongatus caerulescens have many free neuromasts on their body surface. This study examined the ability of the willow shiner larvae to feed on zooplankton by mechanoreception by the free neuromasts. Feeding experiments using untreated larvae and larvae treated with streptomycin, which impairs free neuromast function, were conducted under light and completely dark conditions. The larvae were put into Petri dishes, then nauplii of Artemia salina were introduced. The average number of Artemia eaten by the larvae was expressed as the average ingestion rate of Artemia /10 min. The ingestion rate of Artemia for untreated larvae was 12.3 under light conditions and 10.6 Artemia /10 min. even in complete darkness. The ingestion rate in the larvae treated with streptomycin was 11 under light conditions and only 0.8 Artemia /10 min under dark conditions. The low rate in the treated larvae under dark conditions must be due to impairment by the streptomycin. Therefore, the high ingestion rate for the untreated larvae under dark conditions would be surely dependent on mechanoreception. The results indicate that larval willow shiner is able to feed on zooplankton under limited light conditions.     

小殖盘吸虫和长菲策吸虫体被超微结构

本文为小殖盘吸虫和长菲策吸虫体被的两种电镜观察。两虫前端的口吸盘和生殖吸盘或腹袋孔间有大量无纤毛圆锥型乳突。小殖盘吸虫和长菲策吸虫体表无棘,皮层较厚,分别为13.33～19.33和16.05～19.33μm。皮层含近百列分泌小体,但无线粒体。长菲策吸虫腹袋皮层薄,为2.71～3.21μm。文中对同盘吸虫皮层超微结构特点和反刍兽瘤胃内环境进行了讨论。     

鱼腥藻PCC7120基因asr0757/alr0758的初步研究

为研究鱼腥藻PCC7120(Anabaena sp.)基因asr0757/alr0758在毒素-抗毒素系统中的相关生物学功能,设计了特异性引物,扩增目的片段asr0757和alr0758,将目的基因与p MD18-T载体连接构建克隆载体,并对其进行XhoⅠ和NdeⅠ双酶切,再与表达载体p ET-28a连接构建表达重组菌,重组菌的体外表达在IPTG的诱导下进行。经琼脂糖电泳检测,结果扩增出了大小为210 bp的asr0757和342 bp的alr0758目的基因。经SDS-PAGE电泳检测,表达出相对分子质量分别为8.068 k D和12.534 k D的蛋白质。根据结果可初步认定alr0758为毒素基因,asr0757为抗毒素基因,共同构成鱼腥藻PCC7120毒素-抗毒素系统。     

鱼腥蓝细菌PCC 7120中两个磷酸酶N端结构域活性分析

为了研究鱼腥蓝细菌(A nabaena sp.)PCC 7120中两个PP2C类蛋白磷酸酶PrpJ1和PrpJ2的磷酸酶活性,将编码两个蛋白N端至跨膜区的基因片段重组到质粒中,转化大肠杆菌进行原核表达,获得了可溶性的PrpJ1up和PrpJ2up蛋白.并且以pNPP为底物测定了所得蛋白的磷酸酶活性,双倒数作图法结果显示PrpJ1up蛋白的KK为0.30 mmol/L,Vmax为7.10 nmol/min;PrpJ2up蛋白的Km为0.24 mmol/L,Vmax为0.43 nmol/min.     

鱼腥蓝细菌PCC 7120异形胞的分离

鱼腥蓝细菌PCC 7120是一种光合自养型丝状蓝细菌.缺氮条件下,菌丝上5％～10％的营养细胞分化为异形胞,发挥固氮作用.异形胞是研究生物固氮和细胞分化的理想材料.为研究异形胞生理功能及其发育过程中的基因表达变化,需要从菌丝中分离高质量的异形胞.用2 mg/mL溶菌酶与0.1％ Triton X-100同时处理菌丝30 min,得到了完整性好、纯度高的异形胞.