共查询到19条相似文献，搜索用时 156 毫秒
1.
《广东海洋大学学报》2014,(6)
草鱼呼肠孤病毒(Grass carp reovirus,GCRV)是草鱼出血病的病原。从患病草鱼体内分离到一株草鱼呼肠孤病毒GCRV 096,经反转录PCR,克隆得GCRV 096长度为855 bp的vp7基因,并分析该基因编码蛋白的特性。结果表明,同一基因型分离株VP7蛋白间的差异不大,但不同基因型分离株VP7蛋白间存在很大的差异。对GCRV 096 VP7蛋白生物信息学分析表明,信号肽位点位于20氨基酸处,且VP7含有4个潜在的抗原决定簇。构建GCRV 096 vp7基因的酵母表达载体p GBKT7-S10,并成功转化至酵母中。 相似文献
2.
《广东海洋大学学报》2015,(4)
克隆得一株新分离病毒草鱼呼肠孤病毒(GCRV)096的长为1 981 bp的vp4基因,分析该基因的生物信息及其编码蛋白的特性。生物信息学分析表明,同一基因型GCRV分离株vp4基因间的差异不大,但不同基因型GCRV分离株vp4基因间却存在很大的差异。GCRV 096 VP4蛋白含有多个功能位点、2个跨膜结构域及多个潜在的抗原决定簇。构建p EGFP-N3-VP4真核表达质粒,并转染至草鱼肾(Ctenopharyngodon idellus kidney,CIK)细胞中,经荧光显微镜观察和Western blot鉴定,表明GCRV 096 VP4融合蛋白在CIK细胞中得以成功表达。 相似文献
3.
《广东海洋大学学报》2020,(4)
【目的】研究草鱼呼肠孤病毒(grass carp reovirus, GCRV)096 vp7基因的原核和真核表达及其编码蛋白的免疫原性。【方法】应用RT-PCR技术获得GCRV 096 vp7基因,构建GCRV 096 vp7基因原核表达载体pET-VP7,用原核表达的GCRV 096 VP7蛋白免疫草鱼(Ctenopharyngodon idellus)15 d后,用GCRV 096和GCRV GD108分离株对其攻毒,检测GCRV 096 VP7的免疫原性。构建GCRV 096 vp7基因真核表达载体p EGFP-N3-VP7,分析其在草鱼CIK细胞中的表达。【结果与结论】GCRV 096 vp7基因的开放阅读框ORF (GenBank登录号为JN206665)为831 bp,编码276个氨基酸。成功构建GCRV 096 vp7基因原核表达载体pET-VP7,并在大肠杆菌BL21中诱导表达成功;最优表达条件是0.2 mm/L IPTG、28℃下表达5 h,通过HisTrap HP柱纯化融合蛋白,Western blot分析结果表明,所表达的蛋白为目的蛋白。经免疫及GCRV攻毒后,GCRV 096(Ⅰ型)免疫组vp7基因的表达水平降低(P 0.05),而GCRV GD108(Ⅱ型)免疫组vp7基因的表达无显著差异,表明GCRV 096 VP7蛋白对GCRV096具有良好的免疫效果,但对GCRVGD108无明显的免疫效果。成功构建GCRV096vp7基因真核表达载体pEGFP-N3-VP7,Western blot分析结果表明,GCRV 096 VP7蛋白在草鱼肾(CIK)细胞中成功表达。 相似文献
4.
根据草鱼呼肠孤病毒(Grass Carp Reovirus,GCRV)的vp5基因设计特异性引物,并扩增其开放阅读框序列(open reading frame,ORF),然后连接至pGBKT7载体上构建酵母双杂交诱饵载体pGBKT7-vp5,转化至酵母菌Y2HGold,并进行PCR鉴定以及自激活性和毒性检测。PCR、酶切以及测序表明,pGBKT7-vp5重组诱饵载体构建成功。自激活性和毒性检测显示,阳性重组菌株pGBKT7-vp5/Y2HGold和阴性对照菌株pGBKT7/Y2HGold在SD/-Trp平板上出现大小相一致的乳白色菌落,且在SD/-Trp/X-α-Gel、SD/-Trp/AbA+/X-α-Gel平板上生长出蓝色菌落,而在SD/-Trp/-Ade/-His无菌落出现。表明重组菌株pGBKT7-vp5/Y2HGold表达的融合蛋白激活了报告基因AUR1-C和MEL1,没有激活报告基因ADE2和HIS3并且对宿主菌无毒性。该重组诱饵载体可用于酵母双杂交系统进一步筛选cDNA文库中与其相互作用的蛋白。 相似文献
5.
P1 T重组质粒上含有口蹄疫病毒 (FMDV)GD10分离株的p1cDNA片段 ,以此为模板 ,用PCR方法扩增其中的VP1基因 ,获得大小约 6 40bp的片段。该片段用BglⅡ和BstEⅡ酶切消化后克隆至表达载体 pCAMBIA130 5 .2 ,转化EcoliTOP10感受态细胞。重组质粒经PCR、酶切及序列分析 ,证实VP1基因处于CaMV35S启动子控制 ,且读码框正确 相似文献
6.
口蹄疫病毒VP1植物表达载体的构建 总被引：1，自引：0，他引：1
P1-T重组质粒上含有口蹄疫病毒(FMDV)GD10分离株的p1 cDNA片段，以此为模板，用PCR方法扩增其中的VP1基因，获得大小约640bp的片段。该片段用BglⅡ和BstEⅡ酶切消化后克隆至表达载体pCAMBIA1305．2，转化Ecoli TOPIO感受态细胞。重组质粒经PCR、酶切及序列分析，证实VP1基因处于CaMV35S启动子控制，且读码框正确。 相似文献
7.
采用微生物分离培养技术和巢式PCR技术相结合方法对2011年采自广西北海南万和古城、钦州茅岭和钦州港、防城港企沙和公车的拟穴青蟹(Scylla paramamosain)携带寄生虫、细菌和病毒状况开展调查.寄生虫检测结果显示,仅钦州茅岭拟穴青蟹鳃组织中发现寄生虫茗荷(儿).细菌性病原体检测结果显示,共有11种病原性细菌存在于健康养殖拟穴青蟹体内,弧菌属细菌性病原出现频率最高,其中副溶血弧菌和溶藻弧菌在调查区域内均有分布,其他菌种则呈现一定的地域特异性.病毒性病原体检测结果显示,白斑综合症病毒仅在钦州茅岭拟穴青蟹检出,阳性率为4.94%;双顺反子病毒于北海古城和钦州茅岭拟穴青蟹检出阳性携带率为3.70%;呼肠孤病毒携带率则较高,北海、防城港以及钦州拟穴青蟹均有检出,阳性率达25.93%.由此可见,10-11月广西区内健康养殖拟穴青蟹携带弧菌和呼肠孤病毒的比例远远高于其他病原体,为拟穴青蟹携带的主要病原体. 相似文献
8.
用RT-PCR方法从1个H5N1亚型禽流感病毒分离株A/Chicken/Guangdong/DH/1997扩增NA基因cDNA片段,将其克隆至pMD18-T载体,获得重组质粒pMD-NA,并对其核苷酸序列进行测定和分析。结果表明,该毒株的NA基因长度为1350bp,编码449个氨基酸,与其它H5N1亚型AIV分离株的核苷酸序列同源性为97.0%～99.4%,氨基酸序列同源性为97.7%～99.1%,提示禽流感病毒NA基因保守性较高。NA基因氨基酸序列的聚类分析表明该毒株与来自香港的A/Pheasant/HK/FY155/01和A/Ch/HK/FY150/01两个分离株处于同一进化枝,亲缘关系较近。 相似文献
9.
《广东海洋大学学报》2016,(1)
对感染白斑综合征病毒(WSSV)的凡纳滨对虾(Litopenaeus vannamei)分别注射0.05和0.10μg/μL的vp28-si RNA,干扰效果实验表明,si RNA最佳浓度为0.10μg/μL。凡纳滨对虾感染WSSV 24 h时注射0.10μg/μL的vp28-si RNA,检测免疫后不同时间点的干扰效果。结果表明:在注射vp28-si RNA 6 h后,实验组与对照组WSSV病毒拷贝数差异有统计学意义(P0.05);在48 h时实验组病毒拷贝数急剧下降,对照组则保持较高水平,可较好干扰WSSV病毒复制,干扰效果持续120 h,对凡纳滨对虾的保护率为40%。凡纳滨对虾用0.10 mg/m L肽聚糖(Peptidoglycan)免疫24 h后感染WSSV,检测6、12、24 h注射vp28-si RNA的保护率。结果显示:6 h注射组的保护率为80%,12 h注射组为63.33%,24 h注射组为56.67%。凡纳滨对虾在肽聚糖和vp28-si RNA作用后,可增加其对WSSV抗感染作用,降低死亡率,干扰时间越早,对对虾的保护率越高。 相似文献
10.
海水养殖生物病毒病是世界性难题,淋巴囊肿病毒中国分离株(LCDV-cn)是牙鲆(Paralichthys olivaceus)淋巴囊肿病的病原。以mcp基因为靶基因,设计siRNA-mcp1、siRNA-mcp2、siRNA-mcp3等3条siRNA序列,研究RNA干扰(RNAi)对淋巴囊肿病毒在牙鲆体内复制的影响。结果表明,牙鲆体内注入siRNA-mcp1和siRNA-mcp3后,mcp基因的表达有所下调;统计分析表明,注入siRNA-mcp1后,mcp基因的表达下调明显,具有显著性差异(p<0.05),注入siRNA-mcp2、siRNA-mcp3后,mcp基因的表达下调不明显,差异不显著(p>0.05)。本实验中初步实现了RNAi对LCDV-cn复制的抑制作用。 相似文献
11.
The intestinal bacteria of vertebrates form a close relationship with their host.External and internal conditions of the host,including its habitat,affect the intestinal bacterial community.Similarly,the intestinal bacterial community can,in turn,influence the host,particularly with respect to disease resistance.We compared the intestinal bacterial communities of grass carp that were collected from farm-ponds or a lake.We conducted denaturing gradient gel electrophoresis of amplified 16S rRNA genes,from which 66 different operational taxonomic units were identified.Using both the unweighted pair-group method with arithmetic means clustering and principal component analysis ordination,we found that the intestinal bacterial communities from the two groups of pond fish were clustered together and inset into the clusters of wild fish,except for DF-7,and there was no significant correlation between genetic diversity of grass carp and their intestinal bacterial communities(Mantel one-tailed test,R=0.157,P=0.175).Cetobacterium appeared more frequently in the intestine of grass carp collected from pond.A more thorough understanding of the role played by intestinal microbiota on fish health would be of considerable benefit to the aquaculture industry. 相似文献
12.
The myosin heavy chain (MyHC) is one of the major structural and contracting proteins of muscle. We have isolated the cDNA
clone encoding MyHC of the grass carp, Ctenopharyngodon idella. The sequence comprises 5 934 bp, including a 5 814 bp open reading frame encoding an amino acid sequence of 1 937 residues.
The deduced amino acid sequence showed 69% homology to rabbit fast skeletal MyHC and 73%–76% homology to the MyHCs from the
mandarin fish, walleye pollack, white croaker, chum salmon, and carp. The putative sequences of subfragment-1 and the light
meromyosin region showed 61.4%–80% homology to the corresponding regions of other fish MyHCs. The tissue-specific and developmental
stage-specific expressions of the MyHC gene were analyzed by quantitative real-time PCR. The MyHC gene showed the highest
expression in the muscles compared with the kidney, spleen and intestine. Developmentally, there was a gradual increase in
MyHC mRNA expression from the neural formation stage to the tail bud stage. The highest expression was detected in hatching
larva. Our work on the MyHC gene from the grass carp has provided useful information for fish molecular biology and fish genomics. 相似文献
13.
对网围养殖区内草鱼的活动声波进行了采集与分析,并将采集的声波进行了回放。结果表明,草鱼网围养殖区声波频率范围为0~3.2 kHz,整个频段内没有宽带波峰。在草鱼饥饿、食饵和饱食的不同状态下养殖区内的水声特性有一定的差异。研究表明在网围养殖区通过声波回放能对草鱼起到诱集作用,在投饵回放、投饵不回放及不投饵回放声波的情况下,草鱼的响应时间分别为2.55、3.504、.80 min,存在显著差异。回放持续性声波和间隔性声波在响应时间上不存在显著差异。 相似文献
14.
孙军 《中国海洋湖沼学报》2011,29(1):162-166
The efficacy of grass carp Ctenopharyngodon idella (Cyprinidae) and weevils Neochetina spp. (Curculionidae) to control the aquatic weed, water hyacinth, is investigated in a square net cage (happas) setting at a farm in Cuddalore District, South India. This novel combination of insects and fish is found to be superior to individual treatments for controlling the weed growth within 110 d. The biomass of the weed, number of plants, percentage of flowered plants and chlorophyll contents were studied. The weed biomass is reduced from 5 kg (day 1) to 0.33 kg (day 110) when exposed to grass carp and weevils. The number of plants is reduced to 0.75 in grass carp and weevil exposed happas, while it is 741.5 in the control. The mean number of leaves per plant is also reduced. In addition, the chlorophyll a and b are significantly reduced in happas exposed to the combination of fish and insects when compared to the other treatments. Based on the results of this study, we consider the combined use of grass carp and weevils to be more efficient and sustainable for managing water hyacinths than the use of these organisms individually. 相似文献
15.
6个不同鲤群体的形态差异分析 总被引：4，自引：0，他引：4
对黄河鲤、荷包红鲤、高背荷包红鲤、兴国红鲤、建鲤和黑龙江野鲤等6个鲤群体的12个形态比例性状进行单因素方差分析、主成分分析和聚类分析。单因素方差分析结果表明,除眼后头长/头长外,各鲤群体间在其他比例性状上出现较明显的差异。主成分分析构建了3个主成分,其贡献率分别为35.316%、23.221%、10.146%,累计贡献率为68.683%,并明显可将6个鲤群体划分为2个簇,荷包红鲤和高背荷包红鲤明显与其他群体区分开。聚类分析结果与主成分分析一致。在体高/体长、头长/体长、体厚/体长和尾柄高/尾柄长等4个比例性状上,有些群体的差异系数大于1.28,说明这些群体在这4个指标上的差异可达到亚种水平。结果表明,6个鲤群体在形态上存在一定差异和分化,主要体现在体型和头部特征上。 相似文献
16.
Bothriocephalus acheilognathi is a potentially serious pathogen in wild or cultured fish in worldwide distribution. We examined 58-farmed grass carp from
Nanchang in the Changjiang (Yangtze) River drainage, from which 20.7% were found to harbor the parasite with an infection
intensity of 36.9±54.7. The parasites were identified based on morphology and rDNA ITS sequence analysis. The present report
represents the first record of the parasite in grass carp Ctenopharyngodon idella in the river drainage. 相似文献
17.
EXPERIMENTAL STUDIES ON THE ROLE OF PLANKTIVOROUS FISHES IN THE ELIMINATION OF MICROCYSTIS BLOOM FROM DONGHU LAKE USING ENCLOSURE METHOD 总被引：11，自引：0，他引：11
谢平 《中国海洋湖沼学报》1996,(3)
The hypertrophic subtropic Donghu Lake's dense water bloom(of mainly Microcystis Anaabaena andOscillatoria)that occurred annually from the beginning of the 1970s, has disappeared since 1985. The infiuenceof planktivorous fishes (silver and bighead carps) on the water bloom was studied for three years usingthe enclosure method. The enclosures stocked densely with bighead and/or silver carp were free of waterbloom during the experimental period. The water bloom that appeared in the fish-free enclosures was completelyeliminated in 10-20 days by introduction of silver and/or bighead carp(grass carp was not effectivein controlling water bloom).This study showed clearly that grazing pressure by planktivorous fishes is a keyfactor in eliminating water bloom from the lake. 相似文献
18.
Viral entry into the host is the earliest stage of infection in the viral life cycle in which attachment proteins play a key role. VP31 (WSV340/WSSV396), an envelope protein of white spot syndrome virus (WSSV), contains an Arg-Gly-Asp (RGD) peptide domain known as a cellular attachment site. At present, the process of VP31 interacting with shrimp host cells has not been explored. Therefore, the VP31 gene was cloned into pET30a (+), expressed in Escherichia coli strain BL21 and purified with immobilized metal ion affinity chromatography. Four gill cellular proteins of shrimp (Fenneropenaeus chinensis) were pulled down by an affinity column coupled with recombinant VP31 (rVP31), and the amino acid sequences were identified with MALDI-TOF/TOF mass spectrometry. Hemocyanin, beta-actin, arginine kinase (AK), and an unknown protein were suggested as the putative VP31 receptor proteins. SDS-PAGE showed that AK is the predominant binding protein of VP31. An i n vitro binding activity experiment indicated that recombinant AK’s (rAK) binding activity with rVP31 is comparable to that with the same amount of WSSV. These results suggested that AK, as a member of the phosphagen kinase family, plays a role in WSSV infection. This is the first evidence showing that AK is a binding protein of VP31. Further studies on this topic will elucidate WSSV infection mechanism in the future. 相似文献
19.
The present study preliminarily examined the differences in maximum handling size, prey size and species selectivity of growth hormone transgenic and non-transgenic common carp Cyprinus carpio when foraging on four gastropods species( Bellamya aeruginosa, Radix auricularia, Parafossarulus sinensis and Alocinma longicornis) under laboratory conditions. In the maximum handling size trial, five fish from each age group(1-year-old and 2-year-old) and each genotype(transgenic and non-transgenic) of common carp were individually allowed to feed on B. aeruginosa with wide shell height range. The results showed that maximum handling size increased linearly with fish length, and there was no significant difference in maximum handling size between the two genotypes. In the size selection trial, three pairs of 2-year-old transgenic and non-transgenic carp were individually allowed to feed on three size groups of B. aeruginosa. The results show that the two genotypes of C. carpio favored the small-sized group over the large-sized group. In the species selection trial, three pairs of 2-year-old transgenic and non-transgenic carp were individually allowed to feed on thin-shelled B. aeruginosa and thick-shelled R. auricularia, and five pairs of 2-year-old transgenic and non-transgenic carp were individually allowed to feed on two gastropods species( P. sinensis and A. longicornis) with similar size and shell strength. The results showed that both genotypes preferred thin-shelled Radix auricularia rather than thick-shelled B. aeruginosa, but there were no significant difference in selectivity between the two genotypes when fed on P. sinensis and A. longicornis. The present study indicates that transgenic and non-transgenic C. carpio show similar selectivity of predation on the size-and species-limited gastropods. While this information may be useful for assessing the environmental risk of transgenic carp, it does not necessarily demonstrate that transgenic common carp might have lesser environmental impacts than non-transgenic carp. 相似文献