共查询到20条相似文献，搜索用时 93 毫秒
1.
To improve the expression efficiency of exogenous genes in Chlamydomonas reinhardtii,a high efficient expression vector was constructed.Green fluorescent protein(GFP) was expressed in C.reinhardtii under the control of promoters:RBCS2 and HSP70A-RBCS2.Efficiency of transformation and expression were compared between two transgenic algae:RBCS2 mediated strain Tran-Ⅰ and HSP70A-RBCS2 mediated strain Tran-Ⅱ.Results show that HSP70A-RBCS2 could improve greatly the transformation efficiency by approximately eightfold of RBCS2,and the expression efficiency of GFP in Tran-Ⅱ was at least double of that in Tran-Ⅰ.In addition,a threefold increase of GFP in Tran-Ⅱwas induced by heat shock at 40°C.All of the results demonstrated that HSP70A-RBCS2 was more efficient than RBCS2 in expressing exogenous gene in C.reinhardtii. 相似文献
2.
LU Yongzhong ZHANG Xuecheng 《中国海洋大学学报(英文版)》2007,6(3):244-248
In order to investigate the regulation mechanism of the phycocyanin gene,a series of functional analyses of the upstream sequence of cpcB gene from Arthrospira platensis were conducted in E. coli with green fluorescent protein encoding gene(gfp) as the reporter. Results showed that the gfp gene could express at a high level under the promotion of the upstream sequence,suggesting the existence of some strong promoter elements in it. The expression of GFP was influenced by temperature. Higher temperature led to higher expression level. The bioinformatics analyses followed by mutation analyses on the secondary structure of translation ini-tiation region(TIR) revealed that RNA thermosensor might account for the temperature regulation. 相似文献
3.
To improve the expression efficiency of exogenous genes in Chlamydomonas reinhardtii, a high efficient expression vector was constructed. Green fluorescent protein （GFP） was expressed in C. reinhardtii under the control of promoters： RBCS2 and HSP70A-RBCS2. Efficiency of transformation and expression were compared between two transgenic algae： RBCS2 mediated strain Tran-Ⅰ and HSP70A-RBCS2 mediated strain Tran-Ⅱ. Results show that HSP70A-RBCS2 could improve greatly the transformation efficiency by approximately eightfold of RBCS2, and the expression efficiency of GFP in Tran-Ⅱ was at least double of that in Tran-Ⅰ. In addition, a threefold increase of GFP in Tran-Ⅱ was induced by heat shock at 40℃. All of the results demonstrated that HSP70A-RBCS2 was more efficient than RBCS2 in expressing exogenous gene in C. reinhardtii. 相似文献
4.
A large yellow croaker,Pseudosciaena crocea,spleen(LYCS)cell line was established and the feasibility of using it for foreign gene transfection was evaluaed in this study.Primary culture of LYCS cells was initiated from spleen tissue pieces,which were cultured at 25℃ in Dulbecco’s modiced Eagle medium/F12 medium(DMEM/F12,1:1)(pH7.2),supplemented with 20% fetal bovine serum,carboxymethyl chitosan,chondroitin sulfate,basic fibroblast growth factor(bFGF)and insulin-like growth factor-I(IGF-I).The cultured LYCS cells,in fibroblast shape,proliferated to 100% confluency 20 days later.Chromosome analyses indicated that the LYCS cells exhibited chromosomal aneuploidy with a modal chromosome number of 48 which displayed the normal diploid karyotype of P.crocea(6m+6sm+36t,NF=60).A LYCS cell line,with a population doubling time of 48.7 h at passage 60,has been established and subcultured to passage 70.Transgenic feasibility test demonstrated that positive green fluorescence protein(GFP)expression was observed in LYCS cells after pcDNA3.1-GFP plasmid transfection.In conclusion,a continuous foreign gene trans-fection feasible LYCS cell line has been established successfully.The cell line might serve as a valuable tool for studies of transgenic breeding and has potential applications for different kinds of cytotechnological studies. 相似文献
5.
《中国海洋大学学报(英文版)》2021,(5)
STAT3 plays a particularly important role in several cancer-related signal transduction pathways. Silencing STAT3 via RNA interference or small molecule inhibitors induces the apoptosis of tumor cells, thereby inhibiting the growth of the tumors. In this study, short-hairpin RNA sequences targeting the STAT3 genes were designed, synthesized, and then connected to pGPU6/GFP/Neo plasmids as the shRNA-expression vectors. The expression of STAT3-shRNA was analyzed by real-time PCR, western blotting, and cell apoptosis assay to study the growth and apoptosis of the cells. Then, the effect of STAT3 knockdown on the NCI-H1650 cells was studied in a tumor mouse model. The results revealed that, after an in vitro transfection, the proliferation of NCI-H1650 cells was inhibited, and the cells were induced to apoptosis. The mRNA and protein expression levels of STAT3 were downregulated in the STAT3-sh RNA group. In vivo, the tumor mass and volume in the STAT3-sh RNA group were significantly lower than in the other two groups. Both the in vivo and in vitro results demonstrated a long-period inhibiting effect on NSCLC, especially in vivo,when the tumor inhibition rate could reach 50% in the STAT3-shRNA group, which is an exciting outcome. Moreover, the study of the conjugation of STAT3-shRNA and chitosan-based vectors revealed that they could be combined steadily with good cytocompatibility and transfection efficiency. These results together provide convincing evidence for the application of STAT3-shRNA used in the treatment of non-small lung cancer, which could be a promoting prospect for the development of gene therapy. 相似文献
6.
A cell line,SHK,was derived from the kidney of spotted halibut Verasper variegates.The cell line was subcultured more than 40 passages in minimum essential medium(MEM)supplemented with fetal bovine serum(FBS)and 10 ng ml-1 basic fibroblast growth factor(bFGF).Cell morphology from primary culture and subculture was observed continuously by microscopy.The SHK cell line consisted predominantly of fibroblast-like cells.The cell line was able to grow between 20℃ and 30℃ with the optimum growth at 24℃ and with a reduced growth between 12℃ and 20℃.The growth rate of the cells increased as the proportion of FBS increased from 10% to 20% at 28℃ with optimum growth at the concentration of 20%.The doubling time of the cells was determined to be 44.8 h.Chromosome analysis revealed that 52% of the SHK cells maintained a normal diploid chromosome number (2n=46).The cells were successfully transfected with green fluorescent protein(GFP)reporter plasmids and the expression of GFP gene in the cells indicated the possible utility of the cells in gene expression studies.The cells were infected by lymphosystis disease virus(LCDV)and found to be susceptible to the virus in cytopathic effect(CPE)observation.The infection was confirmed by PCR and electron microscopy experiments,which proved the existence of the viral particles in the cytoplasm of the virus-infected cells. 相似文献
7.
REN Xueying SUI Zhenghong ZHANG Xuecheng 《中国海洋大学学报(英文版)》2006,5(2):146-150
Glyceraldehyde-3-phosphate dehydrogenase （GAPDH） plays important roles in various cellular processes. A cytosolic GAPDH encoding gene （gpd） of Gracilaria/Gracilariopsis lemaneiformis was cloned and characterized. Deduced amino acid sequence of the enzyme of G. lemaneiformis had high homology with those of seven red algae. The 5＇-untranslated regions of the GAPDHs encoding genes of these red algae varied greatly. GAPDHs of these red algae shared the highly conserved glyceraldehyde 3-phosphate dehydrogenase active site ASCTTNCL. However, such active site of Cyanidium caldarium was different from those of the other six algae at the last two residues （CL to LF）, thus the spatial structure of its GAPDH active center may be different from those of the other six. Phylogenetic analysis indicated that GAPDH of G. lemaneiformis might have undergone an evolution similar to those of Porphyra yezoensis, Chondrus crispus, and Gracilaria verrucosa. C. caldarium had a closer evolutionary relationship with Cyanidioschyzon merolae than with Cyanidium sp. Virtual Northern blot analysis revealed that gpd of G. lemaneiformis expressed constitutively, which suggested that it might be house-keeping and could be adapted as an inner control in gene expression analysis of G. lemaneiformis. 相似文献
8.
Plants possess effective mechanisms to respond quickly to the external environment.Rapid activation of phosphatidylinositol-specific phospholipase C(PLC)enzymes occurs after a stimulus.The PLC in Dunaliella salina play s important roles in growth and stress responses.However,the molecular basis of PLC action in D.salina remains little understood.To gain insight into the potential biological functions of this enzyme,we cloned a phospholipase C gene from D.salina in a previous study,named DsPLC(GenBank No.KF573428).Here,we present the prokaryotic expression,purification,and characterization of the DsPLC gene.The entire coding region of DsPLC was inserted into an expression vector pET32 a,and the DsPLC gene was successfully expressed in Escherichia coli.The DsPLC protein was purified and identified using a polyclonal antibody and western blotting.Expressing DsPLC fused with a green fluorescent protein(GFP)in onion showed that DsPLC-GFP was localized to the intracellular membrane.Quantitative real-time PCR analysis revealed that the relative expression of the DsPLC gene was induced significantly by 3.0-mol/L NaCl at 4 h.Our results support the importance of PLC enzymes in plant defense signaling.This study provides a basis for further functional studies of the DsPLC gene and for additional analysis of the potential roles of PLC enzymes in response to abiotic stress. 相似文献
9.
《中国海洋湖沼学报》2016,(1)
A hydrodynamic model and an aquatic ecology model of Dianshan Lake,Shanghai,were built using a hydrodynamic simulation module and the water quality simulation module of Delft3D,which is an integrated modelling suite offered by Deltares. The simulated water elevation,current velocity,and direction were validated with observed data to ensure the reliability of hydrodynamic model. The seasonal growth of different algae was analyzed with consideration of observed and historical data,as well as simulated results. In 2008,the dominant algae in Dianshan Lake was Bacillariophyta from February to March,while it was Chlorophyta from April to May,and Cyanophyta from July to August. In summer,the biomass of Cyanophyta grew quickly,reaching levels much higher than the peaks of Bacillariophyta and Chlorophyta. Algae blooms primarily occurred in the stagnation regions. This phenomenon indicates that water residence time can influence algal growth significantly. A longer water residence time was associated with higher algal growth. Two conclusions were drawn from several simulations: reducing the nutrients inflow had little effect on algal blooms in Dianshan Lake; however,increasing the discharge into Dianshan Lake could change the flow field characteristic and narrow the range of stagnation regions,resulting in inhibition of algal aggregation and propagation and a subsequent reduction in areas of high concentration algae. 相似文献
10.
比较不同温度、光照及海水相对密度条件下无菌与自然带菌波吉卵囊藻(Oocystis borgei)生长的差异。结果表明:在20℃时,无菌藻的比增长率(μ)和叶绿素a(Chla)含量分别为0.398d-1和0.117mg/L,均显著高于带菌藻(P<0.05),在28和35℃时,无菌藻和带菌藻生长无显著性差异(P>0.05);在照度为16μmol·m-2·s-1时,无菌藻平均比增长率μ和Chla含量分别为0.345d-1和0.123mg/L,均显著高于带菌藻(P<0.05),在照度为39和88μmol·m-2·s-1条件下,无菌藻和带菌藻生长无显著性差异(P>0.05);在海水密度为1.007、1.020和1.030g/mL时,无菌藻和带菌藻的生长均无显著性差异(P>0.05),μ和Chla含量分别为0.325～0.374d-1和0.085～0.133mg/L。 相似文献
11.
GONG Qianhong) YU Wengong) * DAI Jixun) LIU Hongquan) ) XU Rifu) GUAN Huashi) and PAN Kehou) ) College of Medicine Pharmaceutics Ocean University of China Qingdao P. R. China ) College of Marine Life Sciences Ocean University of China Qingdao P. R. China ) College of Fisheries Ocean University of China Qingdao P. R. China 《中国海洋大学学报(英文版)》2007,6(1):21-25
1 Introduction Porphyra yezoensis is one of the most widely con-sumed and cultivated seaweeds in the world. Over the past decades, much effort has been made in improving the cultured stocks and developing breeding techniques of alga. At present, nuclear transformation systems of eu-karyotic algae such as green unicellular alga Chlamydo-monas reinhardii (Rochaix and Dillewijn, 1982), diatom Phseodactylum tricornutum (Lioudmila et al., 2000) and simple multicellular green alga Volvox carteri (… 相似文献
12.
Sargassum horneri is a macroalga widespread in North Asia-Pacific region, and these years its bloom has caused huge damage to the environment and the economic in China. To make up the blank on genetic engineering research, a transient transformation system for the multicellular marine brown alga S. horneri was established in this research. The algae used in this research were collected from the Yellow Sea of China and verified as a same species S. horneri with analysis of molecular markers. The S. horneri parietal leaves were transformed with the enhanced green fluorescent gene as the reporter by micro-particle bombardment. The results show that green fluorescent protein(GFP) is an ef fective transgene reporter for S.horneri and that particle bombardment is a suitable method for transformation of S. horneri. Through selection of four dif ferent promoters for EGFP and six groups' bombardment characters, the highest transformation e ciency approximately 1.31% was got with the vector pEGFP-N1 at bombardment characters 900 spi and6 cm distance. This research paves a way for the further research and application of S. horneri. 相似文献
13.
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
14.
Gracilaria tenuistipitata var Liui were mono-cultivated and co-cultivated with Pinctada martensii under high(33) and low(21) salinity conditions in laboratory.The daily growth rate of the alga was determined.Tissue carbon and nitrogen contents,the yield and fractional composition of agar were analyzed.Results showed that:1.Gracilaria grew better under low salinity conditions,the daily growth rate was twice that under high salinity conditions.Co-cultivated algae grew faster than mono-culti-vated algae under low salinity conditions,the daily growth rate was about 37.6?higher.2.Compared with mono-cultivated algae,tissue nitrogen contents of co-cultivated algae were higher,while the C:N ratios were much lower.3.The agar yields of co-cultivated algae were much lower than those of mono-culti-vated algae.Agar yield was found to be negatively correlated to the tissue nitrogen contents,and positively correlated to the C:N ratios.4.The highest fractional yields obtained from co-cultivated algae were extracted with 40?ethanol,while from mono-cultivated algae,the highest fractional yields obtained were extracted with distilled water at room temperature. 相似文献
15.
ＩＮＴＲＯＤＵＣＴＩＯＮＩｎｒｅｃｅｎｔｙｅａｒｓ,ｃｏ ｃｕｌｔｕｒｅ (ｉｎｔｅｇｒａｔｅｄｃｕｌｔｕｒｅ )ｏｆｍａｒｉｎｅｓｐｅｃｉｅｓ (ａｌｇａｅａｎｄａｎｉｍａｌｓ)ｈａｓｒｅｃｅｉｖｅｄｇｒｅａｔａｔｔｅｎｔｉｏｎ .Ｉｎｃｏ ｃｕｌｔｕｒｅｓｙｓｔｅｍｔｈｅａｌｇａｅａｃｔｉｖｅｌｙｕｐｔａｋｅＣＯ2 ａｎｄｔｈｅｍｅｔａｂｏｌｉｃｗａｓｔｅｓ(ｓｕｃｈａｓＮＨ4 +,ＮＯ2 - ,ＮＯ3- ｅｔｃ)ａｓｎｕｔｒｉｅｎｔｓｏｕｒｃｅｓ,ａｎｄｒｅｌｅａｓｅＯ2 ｔｏｔｈｅｓｕｒｒｏｕｎｄｉｎｇｅｎｖｉｒｏｎ … 相似文献
16.
Effects of salinity and nutrients on the growth and chlorophyll fluorescence of Caulerpa lentillifera 总被引：1，自引：0，他引：1
Caulerpa lentillifera is a green algae that distributes worldwide and is cultivated for food. We assessed vegetative propagation of C. lentillifera by measuring the specific growth rate(SGR) and chlorophyll fluorescence of the green algae cultured at different salinities and nutrient levels. The results indicated that C. lentillifera can survive in salinities ranging from 20 to 50,and can develop at salinities of 30 to 40. The maximum SGR for C. lentillifera occurred at a salinity of 35. Both chlorophyll content and the ratio of variable to maximum fluorescence( Fv / Fm) were also at a maximum at a salinity of 35. Photosynthesis was inhibited in salinities greater than 45 and less than 25. Both the maximum SGR and maximum chlorophyll content were found in algae treated with a concentration of 0.5 mmol/L of NO3-N and 0.1 mmol/L of PO 4-P. The photosynthetic capacity of photosystem II(PSII) was inhibited in cultures of C. lentillifera at high nutrient levels. This occurred when NO3-N concentrations were greater than 1.0 mmol/L and when PO4- P concentrations were at 0.4 mmol/L. As there is strong need for large-scale cultivation of C. lentillifera,these data contribute important information to ensure optimal results. 相似文献
17.
A genetic transformation model for the seaweedLaminaria japonica mainly includes the following aspects:
-
1.
The method to introduce foreign genes into the kelp,L. japonica
Biolistic bombardment has been proved to be an effective method to bombard foreign DNA through cell walls into intact cells of both sporophytes and gametophytes. The expression ofcat andlacZ was detected in regenerated sporophytes, which suggests that this method could induce random integration of foreign genes.
Promoters to drive gene expression
-
2.
The CaMV35S promoter was first used by us to induce the expression of GUS gene in brown algae. But results of further studies suggested that CaMV35S could be a tissue-specific promoter. Our use of SV40 promoter resulted in both transient and stable expression oflacZ andcat in sporophytes or gametophytes. No GUS or LacZ background was found in either sporophytes or gametophytes.The regeneration route of transgenic kelp
The regeneration efficiency of explants is still very low. By using female gametophytes as gene hosts and parthenogenesis as regeneration route, CAT activity and LacZ activity were detected in regenerated sporophytes of parthenogenetic kelp. li]4.|The way to select transgenic kelp
-
1.
Results of sensitivity tests showed that kelp was only sensitive to chloramphenicol and hygromycin among many antibiotics. The regenerated sporophytes by parthenogenesis were more sensitive to hygromycin than to chloramphenicol. Resistant kelp was created by transforming female gametophytes with pSV40-CAT and stimulating parthenogenesis followed by selection in medium with lethal concentration of chloramphenicol.
Safety consideration of transgenic kelp
L. japonica was originally introduced from Japan. In China it is a cultured population. The possibility of its negative impact on natural populations is very low. 2) The vectors and target genes used for transformation should be restricted in order to avoid any negative impacts on human health and environment. 3) Specially devised containers (3.6 L, made of 200 μm membrane) were used to ensure that the kelp cannot escape or be eaten by marine animals. 4) To avoid the release of spores, it is very necessary to harvest the kelp at suitable age before the sporangium forms.
18.
XIE Zhihao） ） XIAO Hui） TANG Xuexi） CAI Hengjiang） ） Marine Ecology Laboratory Ocean University of China Qingdao P. R. China ） Faculty of Life Science Biotechnology Ningbo University Ningbo P. R. China 《中国海洋大学学报(英文版)》2009,8(2)
The interspecific interactions between the rotifer Brachionus plicatilis and two harmful algal blooms(HAB) species were investigated experimentally by single culture method. B. plicatilis population and the growth of the two algae were compared at different algal cell densities. The results demonstrated that the B. plicatilis obtained sufficient nutrition from Prorocentrum donghaiense to support net population increase. With exposure to 2.5×104 cells mL-1 of P. donghaiense,the number of B. plicatilis increa... 相似文献
19.
Harmful algal blooms in eutrophic waters pose a serious threat to freshwater ecosystems and human health. In-situ light availability control is one of the most commonly used technologies to suppress algae in lakes and reservoirs. To develop a better understanding of the effects of light on algal growth, specific density, colony size and sinking loss, Anabaena flos-aquae(cyanobacteria) and Scenedesmus obliquus(green algae) were evaluated in varying light scenarios. The results showed that the specific density and colony size of these two species varied during growth, and there were obvious differences among the light scenarios. At the end of exponential phase, S. obliquus incubated under light-limited condition maintained a higher specific density and formed larger aggregates, whereas A. flos-aquae formed a longer filament length. Both species exhibited higher sinking loss rates with lower light availability. These results implied that the sinking loss rate was not always constant but should be considered as a variable response to the change of light availability, and in-situ light availability control might result in a significant increase of the sinking loss of algae due to the change of size and specific density, thereby further affecting the algal biomass in the water column. 相似文献
20.
According to palaeoclimatic and modern instrumental data, numerous studies have indicated that the Arctic climate has undergone a significant warming during the past 100 years, and this may lead to significant impact on the fragile lake ecosystem. In this study, we collected a lake sediment core from the Ny-(A)lesund of Svalbard and determined the concentrations of four pigments including chlorophyll derivatives, total carotenoids, oscillaxanthin and myxoxanthophyll in the sediments. Combined with other physical and chemical proxies such as calcium carbonate, total organic carbon, biogenic silicon etc., we have reconstructed the historical changes of lacustrine primary productivity in Ny-(A)lesund, especially for the past 100 years. The results showed that during the period of Little Ice Age (LIA), the climate was unfavorable for the growth of the lake algae, and thus the lacustrine productivity declined. This result was supported by the relatively low contents of pigment and biogenic silica in the sediments. In contrast, the contents of total organic carbon (TOC) and sediment pigments increased significantly in the upper 5 cm (~1890AD), reflecting the rapid growth of the lake algae, thus the great increase of lacustrine primary productivity, corresponding to the warming climate after LIA. However, the biogenic silica in the upper sediments still had a relatively low level, and this might be related to the growth competition with other algae species. Over the past 100 years, the ratio of Osc/Myx in the sediments decreased continuously, indicative of durative increase of myxoxanthophyll in blue-green algal pigments, and this might imply that the human activity had enhanced the nutrition level of the lake in the Arctic region. 相似文献