76results about How to "Improve transgenic efficiency" patented technology

The invention discloses a white birch transgenosis method, belongs to the field of plant gene engineering and aims at solving the problem that white birch transgenosis is low in efficiency, difficult in sterilization and long in period. The method includes: 1, selecting white birch seeds, soaking, and picking mature seeds which are fully expanded by absorbing, submerged at the bottom of a bottle and germinated completely as transgenosis receptors; 2, sterilizing; 3, preparing engineering bacterium liquid used for infection of the seed receptors; 4, longitudinally cutting the seeds, and dip-dyeing; 5, placing the seeds in a liquid callus induction medium for co-culture; 6, obtaining a transgenosis strain to complete through callus culture, differential culture, subculture and rooting culture. By using the method, culture of a white birch explant is not needed, so that transgenosis period is shortened; a liquid co-culture mode is adopted, so that repeated sterilization steps are omitted, and white birch transgenosis efficiency is improved; conversion efficiency is high, period is short, and the problem that white birch transgenosis is low in efficiency, difficult in sterilization and long in period is solved. The white birch stain is high in transplanting survival rate, and the method is simple to operate and easy to master.

The invention discloses a method for acquiring transgene cottons. The method comprises the following steps of: (1) cutting cotyledons, leaf stalks and a part of hypocotyl of cotton cotyledon seedlings and keeping r 1-3 cm of the hypocoty to obtain tissues to be infected at 25-30 DEG C and with the relative humidity 40-60%, and culturing for 2-3 days under dark environment; (2) scratching double surfaces from the top of a stem tip to the base part of the hypocotyl with the cut of 0.8-1.2 mm in depth; (3) dip-dyeing for 15-45 min in solution A which contains a recombination agrobacterium carrying exogenous genes; (4) co-culturing for 2-4 days and then carrying out resistance screening to obtain the resistant cotton seedling; and (5) grafting the resistant cotton seedling on a cotton rootstock to obtain the transgene cotton. The invention improves the conversion efficiency through adjusting the growth states of the stem tip, determining the appropriate depth of the agrobacterium-converted stem tip infection, adding acetosyringone and adjusting the proper pH value. Transgene plants can be quickly obtained by using the method, and the higher transgene efficiency is higher.

The invention provides a high-efficiency recombined adenovirus containing an ndrg2 gene and the pharmic purpose thereof. The recombined adenovirus contains the ndrg2 gene, a Gateway system and a Virapower TM Adenoviral Expression system are utilized in the construction process, and the adenovirus is packed by a recombination method in vitro. After being cloned to an introduction carrier, a target gene can be efficiently and quickly cloned to other carriers of a receptor by specific recombination sites on the carriers and recombinase without depending on a restriction enzyme. The package of the adenovirus in vitro simplifies the experiment steps, shortens the operation time, and has high multiplication efficiency, high transgene efficiency and high virus yield efficiency as well as the transduction efficiency as high as 100 percent. The invention solves the technical problem of recombination and package of the ndrg2 gene in vitro as well as probes and determines the lethal effect of the high-efficiency recombined adenovirus to brain glioma cells and cells of lung cancer, liver cancer, gastric cancer and colon cancer. The high-efficiency recombined adenovirus can be used as a medicament for treating brain glioma, long cancer, liver cancer, gastric cancer and colon cancer.