鲑鱼生长激素cDNA的分子克隆和序列分析

从太平洋切奴克鲑鱼(Pacific Chinook Salmon,Oncorthychus tschawytscha)垂体poly(A)~+ RNA构建cDNA文库。按照鲑鱼生长激素(sGH)部分氨基酸序列合成两个寡聚脱氧核苷酸探针,它们分别与编码第1—7和第166—172氨基酸序列互补。用探针筛查cDNA文库,得到了完整的sGH cDNA克隆。cDNA序列已测定,包括编码210个氨基酸的编码序列。其中含有22个氨基酸的信号肽序列和188个氨基酸的成熟GH序列。该克隆还包括了5'端和3'端非翻译区,分别为72个和438个碱基对长。与Chum鲑鱼比较表明,核酸序列和氨基酸序列的同源性分别为97%和99%。     

彩鲫Ran基因全长cDNA的克隆及其组织表达特异性分析

根据Ran的保守区序列设计简并引物,结合SMART、cDNA合成及RACE-PCR技术,克隆到彩鲫(Carassius auratus color variety)的Ran基因的cDNA全长,其编码区全长为648bp,编码215个氨基酸。采用BIAST程序在NCBI数据库中对其进行同源基因搜索,结果表明,其推测的编码氨基酸序列与斑马鱼和鲑鱼的Ran基因编码的氨基酸序列的同源性分别高达98％和97％。还对其编码区全长序列进行了原核表达,以经纯化的表达蛋白免疫家兔,制备出了具有较高特异性的多克隆抗体。采用Western blotting进行的组织特异性分析表明,Ran蛋白在卵巢、精巢和肾中表达,在心、脑、肝、脾和肌肉中不表达。本工作为采用免疫耗竭法、免疫共沉淀、体外系统模拟等方法进一步研究鱼类．Ran基因的生理功能及分离鉴定其结合蛋白提供了良好基础。     

谷子肌动蛋白基因的克隆及序列分析

以谷子 (Setariaitalica)为材料 ,提取总RNA。根据植物肌动蛋白基因编码区的两端的保守序列设计了简并引物 ,用 5’RACE方法扩增出了谷子肌动蛋白基因编码区序列。以豌豆肌动蛋白cDNA作探针进行的Southern杂交分析表明扩增出了目的基因。将所获得的片段克隆到T载体后进行测序 ,序列分析结果表明 :谷子肌动蛋白基因的编码区长 1 1 3 1个核苷酸 ,编码了 3 77个氨基酸 ;所得序列 (命名为MIAc)与GenBank中注册的肌动蛋白基因序列的相似性均在 6 0 %以上 ,与其它肌动蛋白氨基酸序列的相似性达 89%以上。根据高等植物肌动蛋白序列相似性重建了进化树 ,表明谷子肌动蛋白与水稻肌动蛋白异型体RAc2和RAc3之间的亲缘关系最为密切 ,在进化过程中分化时间最为接近     

谷子肌动蛋白基因的克隆及序列分析

以谷子（Setaria italica）为材料,提取总RNA。根据植物肌动蛋白基因编码区的两端的保守序列设计了简并引物,用5'RACE方法扩增出了谷子肌动蛋白基因编码区序列。以豌豆肌动蛋白cDNA作探针进行的Southern杂交分析表明扩增出了目的基因。将所获得的片段克隆到T载体后进行测序,序列分析结果表明：谷子肌动蛋白基因的编码区长1131个核苷酸,编码了377个氨基酸;所得序列(命名为MIAc)与GenBank中注册的肌动蛋白基因序列的相似性均在60%以上,与其它肌动蛋白氨基酸序列的相似性达89%以上。根据高等植物肌动蛋白序列相似性重建了进化树,表明谷子肌动蛋白与水稻肌动蛋白异型体RAc2和RAc3之间的亲缘关系 最为密切,在进化过程中分化时间最为接近。     

大鼠肝含5~1端氨甲酰磷酸合成酶Ⅰ基因的克隆

为研究氨甲酰磷酸合成酶Ⅰ(CPS Ⅰ)基因表达的调控,我们根据CPS Ⅰ编码1—10氨基酸的mR NA顺序,合成一个30碱基的寡聚核苷酸。以它作为探针从大鼠肝基因库中筛选出一个阳性克隆,λ10。对其中4.5kb插入片断的内切酶谱及部分核苷酸序列测定证明它确含5′上游CPS Ⅰ基因DNA序列。     

玉米中一种新的蛋白激酶电子克隆与序列分析

目的：电子克隆玉米中一种新的蛋白激酶基因。方法：以拟南芥中一个蛋白激酶的氨基酸序列为探针,对玉米的EST数据库进行同源性检索和筛选并克隆。结果：序列分析显示该cDNA长1121bp,有一个450bp的开放阅读框,编码149个氨基酸,且具有保守苏氨酸/丝氨酸蛋白激酶结构域和TEY基元,说明所克隆的cDNA序列为玉米的MAPK全长cDNA。结论：所克隆的cDNA序列为玉米的MAPK全长cDNA。     

人胎肝唾液酸转移酶基因的克隆及测序*

根据已克隆的唾液酸转移酶的保守区的序列,以人胎肝mRNA为模板扩增出150bp的片段并测序。其中一个片段(s38)与已克隆的唾液酸转移酶的活性中心有57％～97％的同源性。根据s38的序列合成寡核苷酸并标记后用作探针筛选人胎肝cDNA文库。从文库中分离了一个编码α2,3－唾液酸转移酶的cDNA。该cDNA序列含一个编码340个氨基酸的开放读框,推导的氨基酸序列与人颌下腺(Galβ1,GalNAc。Α2,3－唾液酸转移酶相同,与猪颌下腺α2,3－唾液酸转移酶有83.2％的同源性。表明从人胎肝cDNA文库中分离的cDNA所编码的蛋白为Galβ1,3GalNAcα2,3－唾液酸转移酶。     

天花粉胰蛋白酶抑制剂基因的克隆及DNA序列分析

从天花粉块茎中分离纯化的天花粉胰蛋白酶抑制剂ＴＴＩ是目前已知的最小的蛋白酶抑制剂,它属于南瓜族蛋白酶抑制剂家族;由２７个氨基酸残基组成,含三对二硫键。本文用ＰＣＲ方法扩增一特异探针;结合传统的筛库方法,从天花粉ｃＤＮＡ基因库中筛选到含ＴＴＩ基因的克隆,经序列测定,得到了ＴＴＩ的ｃＤＮＡ全序列。其读框编码区编码的是一个由６５个氨基酸组成的Ｐｒｅ－Ｐｒｏ－ＴＴＩ,Ｐｒｅ与Ｐｒｏ分别含有２４个和１４个氨基酸。由ｃＤＮＡ序列推论的氨基酸序列和已测定的氨基酸序列完全相同。     

巴西固氮螺菌ntrBC基因的克隆与核苷酸序列分析

以EMBL3为载体,构建了巴西固氮螺菌(Azospirillum brasilense)Yu62的基因文库.以巴西固氮螺菌Yu62中PCR扩增出的450bp DNA 片断作为探针,对该基因文库进行筛选,得到了10个阳性克隆(EA1—EA10),其中含有两种不同类型的克隆,分别以EA4和EA9为代表.对EA4的杂交分析发现目的基因位于2.9kb EcoRI片段上.DNA序列分析结果表明该片段含有完整的ntrC编码区,其编码产物由480个氨基酸组成.分子量为53469;ntrC上游是完整的ntrB编码区,其编码产物由400个氨基酸组成,分子量为43487.对相应的NtrC和NtrB氨基酸序列进行同源性分析,说明巴西固氮螺菌与根瘤菌的亲缘关系较与其它自生固氮菌的更为接近.     

Molecular cloning and expression of salmon prolactin cDNA

Prolactin was purified from chum salmon pituitaries. It was resolved into two variants by reverse-phase high-performance liquid chromatography. A cDNA library was prepared from Pacific chinook salmon pituitaries. Salmon prolactin gene was screened using a synthetic oligonucleotide based on partial protein sequence. A positive clone (PRL-10) was identified and sequenced. It is a full-size clone containing 1.1 kb and coding for a preprolactin of 211 amino acids. A modified prolactin plasmid (PRL-10A), in which the 5' untranslated sequence and the nucleotide sequence coding for the signal peptide of prolactin were deleted, was reconstructed into an expression vector using the heat-inducible lambda pL promotor. Mature prolactin, a single polypeptide of 22 kDa, was efficiently expressed in the bacteria at an elevated temperature.     

The functional relationship between peak spring floods and survival and growth of juvenile Atlantic Salmon (Salmo salar) and Brown Trout (Salmo trutta)

1. The effects of high spring floods on survival and growth of juvenile Atlantic Salmon, Salmo salar, and Brown Trout, Salmo trutta , are explored, using data from a long-term study in the River Saltdalselv, northern Norway. The flow regime in this river is typical for northern rivers.
2. There was considerable variation in year class strength of both species.
3. Mortality of Atlantic Salmon increased significantly in years with high discharge during the alevin stage as well as the first week after emergence. High discharge during the egg stage and more than 1 week after emergence seemed to be of minor importance. Water temperature at emergence was rather high (average 10·5 °C) and did not significantly affect year class strength.
4. Brown Trout emerged earlier than Atlantic Salmon at an average water temperature of 8·2 °C. Highest mortality was observed in years with low water temperatures at emergence as well as high discharge during the alevin stage.
5. For 1-year-old fish or older, the size of the spring peak flood did not influence mortality significantly.
6. Growth of Atlantic Salmon parr was diminished in years with a high peak spring flood. A similar effect on Brown Trout was not detected.     

Occurrence of an adult Atlantic salmon in the River Rhône, France

A large salmonid was caught in the commercial fisheries in the River Rhône (Mediterranean basin, France). Using morphological criteria and DNA analysis, the fish has been identified as an adult Atlantic Salmon Salmo salar .     

Gene expression patterns in Atlantic salmon Salmo salar: gene expression during osmoregulation in intestine tissue

Ireland has the world's largest stocks of wild Atlantic salmon. A better understanding of gene expression will benefit conservation of wild stock as well as salmon aquaculture. We describe the PRTLI project designed to advance the fundamental understanding of the genome of Atlantic Salmon Salmo salar . The major objective is to create the first comprehensive database of gene expression and functional information using cDNA libraries and Microarray technology. One key area of interest to salmon biology is osmoregulation, which is critical to the ability of salmon to adapt in seawater. Tissues implicated in this process are the gills, intestine and skin. To initiate studies, SSH (suppression subtractive hybridization) libraries were constructed from intestine RNA extracted from smolts sampled in January and May. A number of potentially interesting clones have been identified, among those a heat shock protein, hsp90 in the reverse library. Others SSH libraries from various tissues (pituitary, hypothalamus, brain, gill, intestine, head kidney and spleen) have also been constructed and will be used to construct a 5000 clone microarray slide. This slide will then be used to elucidate gene expression profiles in various tissues. Further sample collection has been carried out to answer questions regarding biologicaldifferences between one‐ year and two‐year old parr and wild and hatchery smolt.     

Short‐term response of fish communities and water chemistry to breaching of a causeway in the Sarita River Estuary,British Columbia,Canada

On the west coast of Vancouver Island, Canada, in the Treaty Settlement Lands of the Huu‐ay‐aht First Nation, a causeway isolating the southern portion of the Sarita River estuary was breached in 2018 to enhance juvenile salmon and tidal water access into the southern portion of the estuary. Short‐term goals of breaching were to: (1) enhance juvenile salmon access through the causeway; (2) promote colonization of Chinook and Chum Salmon in pools, while retaining Coho Salmon; (3) monitor the entire estuary fish community, including non‐salmonids, to assess breaching success; and (4) homogenize water chemistry in pool and channel ecosystems currently fragmented by the causeway. Immediately following the breach, both fish community and water chemistry measurements indicate that these goals were achieved. Chinook and Chum Salmon utilization increased inside and outside of the causeway following breaching. Similarly, Chinook and Chum salmon were observed in pool ecosystems, and Coho Salmon were also retained in pools following breaching. Water chemistry was homogenized post‐breach, primarily due to increased saltwater penetration. While continued monitoring is required, short‐term indicators suggest that breaching was effective in increasing connectivity of the Sarita Estuary, allowing access to 1.4 km² of habitat that had previously been inaccessible to juvenile salmon.     

Zooplankton ecology and trophic resources for rearing native fish on an agricultural floodplain in the Yolo Bypass California,USA

Out-migrating juvenile Chinook Salmon Oncorhynchus tshawytscha in California’s Central Valley lack frequent access to historical off-channel habitats such as floodplains. However, many regions have agricultural floodplains that may provide habitat value to young salmon. To determine the suitability of agricultural floodplain, this study tested whether winter-inundated rice fields in a historic flood basin in California’s Central Valley could provide adequate food resources for rearing juvenile Chinook Salmon. We examined the suitability of flooded rice fields for three post-harvest habitat types: stubble, fallow, and disced. Soil emergent and pelagic zooplankton communities were compared to determine colonization sources. Winter-inundated rice fields had high densities of zooplankton, which increased over the course of the study. Daphnia pulex, a large-bodied cladoceran and an excellent forage species of juvenile Chinook Salmon, was abundant in our study. Cladocerans colonized via source water while ostracods likely colonized from a soil egg bank. Overall, there was no discernable effect of habitat type on zooplankton community structure or density, except for D. pulex. Our results suggest that flooded agricultural rearing habitat can support juvenile Chinook Salmon based on high densities of zooplankton and other suitable habitat conditions have the potential to support a robust aquatic food web.     

Benthos and macroinvertebrate drift in six streams differing in alkalinity

The productive capacity of aquatic systems often is equated with the ‘chemical richness’ of the water. A primary objective of the present study was to relate macroinvertebrate benthos and drift to a streams' productive capacity as indicated by absolute levels of alkalinity. We tested this relationship in six 2nd–3rd order tributaries of the Salmon River, Idaho that ranged in alkalinity from 50 to 360 mg 1⁻¹. Benthic density and biomass, drift biomass, and benthic organic matter increased with increasing levels of alkalinity, although not all relationships were significant. The proportion of drift biomass to benthic biomass was similar among study streams suggesting that drift was primarily passive during the study period. The data suggest that spatial variations in landscape-scale geology may indirectly affect spatial patterns of macroinvertebrate benthic and drift standing crops among streams within a single river basin by mediating lotic chemical richness as found among tributaries of the Salmon River basin. Author for correspondence     

Announcement

Rowland, A.P. (Ed,) (1987) Chemical Analysis in Environmental Research.
Shaw, S.A. & Muir, J.F. (1987) Salmon: Economics and Marketing.     

Ontogenetic shifts in the diets of juvenile Chinook Salmon: new insight from stable isotopes and fatty acids

Variations in marine prey availability and nutritional quality can affect juvenile salmon growth and survival during early ocean residence. Salmon growth, and hence survival, may be related to the onset of piscivory, but there is limited knowledge on the interplay between the prey field, environment, and salmon ontogeny. Subyearling Chinook Salmon (Oncorhynchus tshawytscha) and their potential prey were sampled in coastal waters off Willapa Bay, USA to explore this issue. Three seasonal prey assemblages were identified, occurring in spring (May), early summer (June – July), and late summer (August – September). The onset of piscivory, based on salmon stomach contents, fatty acids, and stable isotopes occurred later in 2011 compared to 2012, and coincided with the appearance of Northern Anchovy (Engraulis mordax). Salmon fork length (FL) and carbon isotope values (δ¹³C) increased with a fatty acid biomarker for marine phytoplankton and decreased with a freshwater marker, indicating dietary carbon sources changed as salmon emigrated from the Columbia River. Salmon FL also increased with nitrogen isotope ratios (δ¹⁵N), trophic position, and a fatty acid marker for piscivory – a consequence of the ontogenetic shift in diet to fish. Salmon grew faster and obtained larger size and condition by September 2011 compared to 2012, which was related to inter-annual differences in ocean conditions and the duration over which Northern Anchovy were available. Our results support the idea that juvenile salmon growth depends on the onset and duration of piscivory, suggesting both of these factors may be important components of lifetime growth and fitness.