Disclosed are a construction method and application of a microorganism capable of realizing high production of lacto-N-neotetraose, belonging to the field of microbial genetic engineering. Coding genes of ?-1,3-acetyl glucosamine transferase, ?-1,4-galactosyl transferase and/or UDP-glucose 4 epimerase are over-expressed on the basis of a strain which is previously constructed by the team and is subjected to related-gene knockout, thus enabling the strain to have a synthesis capability of producing the lacto-N-neotetraose. The present disclosure accurately regulates the carbon flux of a metabolic pathway and relieves the metabolic stress by screening the high-efficiency ?-1,4-galactosyl transferase gene and regulating the expression of IgtA, Aa-?-1,4-GalT and galE in a lacto-N-neotetraose synthesis pathway in a combined manner. In a shake flask experiment, the lacto-N-neotetraose production capacity of Escherichia coli is 0.91 g/L. The lacto-N-neotetraose yield in a 3 L fermentation tank reaches 12.14 g/L. Therefore, the microorganism has an industrial application prospect.

The present invention discloses a genetically engineered bacteria, which is E. coli integrated with lysogenic ?DE3, and lacZ gene is completely inactivated, but does not affect exogenous protein expression of the genetically engineered bacteria. The present invention also discloses a method for culturing the genetically engineered bacteria, and a method for preparing human milk oligosaccharides using the same, and use of the genetically engineered bacteria. The genetically engineered bacteria of the present invention can efficiently produce human milk oligosaccharides, such as 2-fucosyllactose, and have wide industrial application prospects.

This patent proves that enucleated red blood cells are the product of biological evolution and disabled red blood cells due to functional defects. Platelets are the nuclei removed during the enucleation of nucleated red blood cells. They are useful components for blood coagulation and hemostasis, but not essential components. Nucleated erythrocytes are fully functional erythrocytes. The preparation of nucleated red blood cell blood, especially the universal blood without blood type, not only has all the functions of red blood cells, but also can be replicated in the blood, improving the physiological performance of blood and reducing the cost and time of artificial blood.

Provided are compositions and methods for effective delivery of polynucleotides into the nuclei of cells through electroporation. A new buffer system is provided that facilitates the delivery with greatly reduced cell toxicity. The buffer may include succinate, mannitol, a sugar, glutamine or an analog, and an antioxidant. Also provided are methods that are particularly suitable for delivering an RNA or protein into a cell by electrophoresis in a solution having an osmotic pressure greater than 310 mOsmol/kg.

A method is disclosed for cryopreservation of cardiocytes derived from pluripotent stem cells or mesenchymal stem cells derived from adipose tissue or bone marrow, the method maintaining the function of the cardiocytes derived from differentiated pluripotent stem cells or mesenchymal stem cells derived from adipose tissue or bone marrow, and yet reducing the possibility for tumorigenesis of undifferentiated pluripotent stem cells or mesenchymal stem cells derived from adipose tissue or bone marrow. A method is also disclosed for cryopreservation of cardiocytes derived from pluripotent stem cells or mesenchymal stem cells (derived from adipose tissue or bone marrow, the method including dissociating cells from a cell population which has been induced to differentiate into cardiocytes from pluripotent stem cells or mesenchymal stem cells derived from adipose tissue or bone marrow.

The present invention refers to a freeze-dried composition consisting of an isolated microorganism belonging to the Mycobacterium tuberucolosis complex, preferably a M. tuberculos clinical isolate, more preferably M. tuberculosis clinical isolate, characterized in that it comprises a PhoP? phenotype by the inactivation by a genetic deletion of the Rv0757 gene and the deletion of a second gene, Rv2930 (fadD26), that prevents PDIM production (PDIM? phenotype) (the MTBVAC strain), and sucrose and sodium glutamate as stabilizers or excipients. The present invention further refers to the reconstituted composition obtained by adding water, preferably sterilized water for injection, to the freeze-dried composition as well as uses thereof, in particular for use as a prophylactic agent to those at risk of infection with M. tuberulosis or those at risk of developing tuberculosis disease, or as secondary agents for treating infected tuberculosis patients.

The invention provides a novel method for controlling and/or treating the growth of biofilms by promoting growth of the periphery cells and/or increasing nutrient consumption by the periphery cells so that the growth of peripheral cells is not only independent from nutrients (e.g., ammonium) provided by the interior cells of the biofilms, but also causes interior cells starved to death. The accessible peripheral cells are then treated and/or eliminated by biofilm control substance of antibiotics or toxic chemicals. Therefore, the invention method controls and eliminates both peripheral and interior cells within biofilms. The invention method can be used in various industries, such as clinical and dental medicine and medical equipment, food and oil industry, and water supply systems.

In certain embodiments, the present invention provides Qualitatively Mapping Amino Acid Properties (QMAP) cell culture media comprising an amino acid component, wherein the amino acids are present in the component at uniform concentrations; an inorganic salt component; an RPMI 1640 Vitamin Solution component; and auxiliary ingredient component. In certain aspects, the present invention also provides methods of making the QMAP cell culture media and using the QMAP cell culture media to culture cells.

The present disclosure relates to an improved method for producing ergothioneine, comprising the steps of: (a) inoculating Pleurotus ostreatus strain CGMCC No. 6232 into a seed medium, and culturing it to prepare a seed liquor, wherein the seed medium uses soybean cake powder as nitrogen source; and (b) inoculating the seed liquor into a fermentation basal medium, and then culturing it to obtain a fermentation broth of Pleurotus ostreatus mycelia. Further, any one or more members selected from NH.sub.4Cl, NH.sub.4NO.sub.3, NaCl, polyethylene glycol, folic acid, vitamin B1 (VB1), indolebutyric acid, citric acid, pyruvic acid, arginine, lysine, leucine, aspartic acid, glutamic acid, betaine, histidine, cysteine, methionine, tween, span, chitosan, Fluconazole, Miconazole, Ketoconazole, ethylenediaminetetraacetic acid (EDTA), isopropyl alcohol and dimethyl sulfoxide are added into the fermentation basal medium.

The present disclosure relates, in general, to a media, e.g., a serum replacement, media supplement, complete media or cryopreservation media, comprising a base physiological buffer and liposomes comprising cholesterol, phosphatidylcholine and fatty acids. It is contemplated that media provides advantages to improve cell growth in culture compared to cells cultured not using the serum replacement described herein.