ABSTRACT
We obtained intergenus somatic hybrid between Alhagi pseudalhagi and Astragalus cicer by using electroporation. Agrobacterium rhizogenes A4-transformed A. pseudalhagi protoplasts were treated with iodoacetamide so that they were unable to sustain divisions. A. cicer protoplasts were isolated from a methionine-resistant mutant and did not survive in the medium without plant growth regulator. The intergenus somatic hybrid cells were selected based on physiological complementation between the two parents. We optimized some parameters of electroporation, such as direct current impulse, alternating current impulse and the impulse number. We identified ten hybrid clones by morphological observation, checking the chromosome numbers, isoenzyme analysis and random amplified polymorphic DNA analysis, and obtained regenerated plantlets from three hybrid clones.
Subject(s)
Astragalus Plant , Genetics , Physiology , Electroporation , Fabaceae , Genetics , Physiology , Hybridization, Genetic , Transformation, GeneticABSTRACT
In this report, the biological activity of the recombinant Arresten expressed in Nicotiana tabacum was studied. The gene coding for the tumor angiogenesis inhibitor Arresten was PCR-amplified from the plasmid pCA and its plant expression vector named pCAMBIAarr was constructed by inserting the Arresten cDNA fragment into the NcoI/BstEII sites of the plant binary expression vector pCAMBIA1301. Then pCAMBIAarr was transferred into Agrobacterium tumefacien LBA4404 by the freeze-thaw method. The adventitious shoots and regenerated plants of Nicotiana tabacum with hygromycinB-resistance were obtained via Agrobacterium-mediated leaf disk transformation method. Southern hybridization, RT-PCR and Western blotting analysis showed that the Arresten cDNA was integrated into the genome of some of the regenerated plants and the recombinant Arresten was expressed with a molecular size of 26 kD. Recombinant Arresten purified from transgenic tobacco leaves had an anti-proliferative effect on bovine endothelial cells. We speculate that biologically active recombinant Arresten can be produced by using plants as bioreactors.
Subject(s)
Agrobacterium tumefaciens , Genetics , Angiogenesis Inhibitors , Genetics , Collagen Type IV , Genetics , Genetic Vectors , Genetics , Plants, Genetically Modified , Genetics , Metabolism , Recombinant Proteins , Genetics , Nicotiana , Genetics , Metabolism , Transformation, GeneticABSTRACT
Purpose:To explore the relationship between the expression of nm23mRNA,p16 and the oncogenesis,development and metastasis of lung cancer.Methods:Expressional level of nm23mRNA,p16 was detected in 72 human lung cancer samples and 23 normal tissues adjacent to cancer respectively,and their correlation with lymph node metastasis and the degree of cell differentiation was studied by immunohistochemistry (IHC) and in situ hybridization (ISH) methods on tissue microarray.Results:The positive expression for nm23mRNA in cancer samples,in human tissues adjacent to cancer and in primary cancer with lymph node metastasis was 55.6%(40/72),82.6%(19/23),25.0%(6/24) respectively.The positive expression for p16 was 45.8%(33/72),73.9%(17/23),25.0%(6/24) respectively.The expression level of nm23mRNA and p16 were closely related to the degree of cell differentiation( P 0.05).Conclusions:These results suggest that nm23 gene and p16 gene are closely related to the degree of cell differentiation and lymph node metastasis of lung cancer.The lower expression level of nm23mRNA and p16 predicts the metastasis of lung cancer, and the two genes may be an index in monitoring the status and evaluating the prognosis of the lung cancer.
ABSTRACT
Object The influence of producing area and plant age on oxymatrine levels in the dried root of Sophora flavescens Ait. has been investigated to provide useful information to optimize the areas for mass scale propagation, proper management and species conservation. Methods The oxymatrine was quantified by HPLC and various statistical treatments were carried out following SPSS 9.0 and Microsoft Excel. Results The oxymatrine concentrations in the root samples collected from 17 provinces and regions differed from 0.494% to 4.127% and the maximum oxymatrine content (3.493%) was recorded in the root sample collected from Heilongjiang Province. Analysis of variance indicates a highly significant difference in the oxymatrine content of roots collected from different provinces, and the samples from the cold arid northern high-latitude areas had higher oxymatrine than those from warm humid southern low-latitude areas in the mainland China. Plant age is positively related to the total root biomass and oxymatrine content. Conclusion The oxymatrine content in the roots of S. flavescens is quite different between different areas and plant ages. The S. flavescens growing in the cold arid northern high-latitude areas had higher oxymatrine concentration.
ABSTRACT
Callus of Gentiana crassicaulis was initiated from hypocotyl and cotyledon on MS medium supplemented with 2mg/L 2.4-D and 0.5 mg/L BA.Induction frequencies of callus were 100%.Somatic embryos were formed on MS medium containing 2mg/L BA,3mg/L ZT, 1mg/L NAA,3mg/L GA,three time MS organics and vitamins,6% sucrose and 500 mg/L LH and three time FeSO4 (Na2EDTA) .40% Callus produced somatic embryos.It developed into complete plants on hormon-free MS medium.