38 results about "Suppression subtractive hybridization" patented technology

The invention discloses a plant high-pH saline-alkaline tolerance gene SucHP and researches the expression of a high-pH saline and alkaline stress induction gene in a common seepweed herb seedling period by using suppression subtractive hybridization and high-density lattice film technology. The plant high-pH saline-alkaline tolerance gene SucHP can be prepared by the following steps of: firstly separating a gene of a coding tonoplast proton pump from common seepweed herbs; constructing a sense expression vector of the gene; and converting Arabidopsis. A transgenic plant has high high-pH saline-alkaline tolerance and can normally grow under the conditions of 100mM NaHCO3+200mM NaCl and 8.5 of pH. The plant high-pH saline-alkaline tolerance gene SucHP can be used for plant genetic transformation and increases plant saline and alkaline resistance.

The invention provides a method for homogenization subtractive hybridization of full length cDNA. The method comprises the following steps in sequence: (1) synthesizing a first chain cDNA; (2) synthesizing a double chain cDNA; (3) selective connection and primer extension; (4) subtractive hybridization; (5) PCR augmentation of cDNA of differential expression; wherein, the PCR products of the subtractive hybridization can be directly cloned to a vector so as to construct cDNA libraries, where differentially expressed genes are concentrated; the libraries can be utilized to analyze gene expression, prepare gene chips and be marked as probe which selects the differentially expressed genes from the existing libraries; in addition, if the subtractive efficiency is not high enough, the subtractive hybridization can repeat for the PCR products of subtractive hybridization to improve the homogenization and the subtractive efficiency. The method for homogenization subtractive hybridization of the full length cDNA (FNSH) creates the beneficial effects of high subtractive efficiency of differential genes, high efficiency of enriching differential expressed genes, high homogenization and gainof the full length cDNA.

The invention provides soybean purple acid phosphatase GmPAP36 and an encoding gene and application thereof. A low-phosphorous induction soybean root system cDNA subtractive library is established based on the suppression subtractive hybridization technique, low-phosphorous stress inducible expression acid phosphatase candidate EST is selected from the subtractive library, and on this basis, an acid phosphatase candidate gene sequence is cloned and the expression difference of a candidate gene under the phytate phosphorus processing condition is analyzed with the real-time quantitative PCR technique; a gene prokaryotic expression vector is established to achieve gene prokaryotic expression; meanwhile, a gene eukaryotic expression vector is established, arabidopsis and soybean conversion is conducted to obtain transgenosis positive plants, the biological function of the gene GmPAP36 is analyzed, and functional genes are provided for efficient transgenosis breeding of plant phosphorus.