Mouse with human nerve growth factor transgenes as well as preparation method and application of mouse

Abstract

The invention discloses a mouse transfected with human nerve growth factor gene and its preparation method and application. Specifically, the present invention relates to the use of transgenic methods to obtain transgenic mice whose salivary glands specifically express human NGF, and the saliva of transgenic mice prepared by this method can secrete human NGF, which can be used for the treatment of human diseases after separation and purification. The method comprises the following steps: (1) constructing a transposon vector, which contains human NGF gene sequence, parotid secretion protein gene promoter sequence, transposon piggyBac 5' and 3' terminal repeat sequence; (2) constructing A good transposon carrier is injected into the male pronucleus of the fertilized mouse egg, and then the injected mouse fertilized egg is transplanted into the uterus of a surrogate mother mouse, and the surrogate mother mouse is raised until the mouse is born; The mice were tested, and the transgenic mice whose human NGF gene could be stably inherited in the offspring were screened out; (4) Human NGF was isolated and purified from the saliva of the mouse transfected with the human nerve growth factor gene.

Description

technical field [0001] The present invention relates to an animal obtained by a transgenic method and its preparation method and application, in particular to a mouse transfected with human nerve growth factor gene and its preparation method and application. Background technique [0002] Nerve growth factor (NerveGrowthFactor, referred to as NGF) is the first member of the neurotrophin family, which was discovered by Cohen and Levi-Montalcini in 1954, and its function was elucidated in subsequent studies. Cohen and Levi-Montalcini received the 1986 Nobel Prize in Medicine or Physiology for this. NGF not only plays an important role in the proliferation and differentiation of nerve cells, but also plays an important role in the growth and activity of non-nerve cells, such as immune cells, fibroblasts, pancreatic cells, and cardiomyocytes. Therefore, NGF has good potential therapeutic effects on both neurological and non-neurological diseases. [0003] Because mouse NGF (mou...

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Problems solved by technology

However, the mammary gland has the following defects as a bioreactor: 1) only sexually mature and lactating female animals can secrete milk; 2) the secretion of milk is limited; 3) milk contains a large amount of endogenous protein, which is not conducive to the production The specifically expressed exogenous protein, namely human NGF, was purified from

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