Salivary gland tissue-specific transgene vector and its construction method

Abstract

The invention discloses a salivary gland tissue specific transgenic carrier and a construction method thereof. The salivary gland tissue specific transgenic carrier is obtained by emerging an upstream regulation region sequence of a mice parotid gland protein promoter and a visual screened neo-EGFP combined double-label to a piggyBac transposon system and a Lox-Cre enzyme system; the transgenic carrier can make protein express in a particular tissue to achieve a high-efficient large-section emerging efficiency. The carrier also has a green luminescence label which is convenient for screening and a neomycin resistance gene which is convenient for eukaryotic cell screening, and makes the screening of transgenic cells and identification of transgenic animals be simpler, more direct, and more accurate. The transgenic carrier provided by the invention is a salivary gland specific expression high-efficient integration universal carrier, and is capable of being applied to fields of transgenic animals such as transgenic mice and pig, and the like. The invention clones the upstream regulation region of FVB mice for the first time, the cloning method is simple and high efficient, and the difficulty in construction of a large section carrier is effectively overcome.

Description

technical field [0001] The invention belongs to the field of biotechnology. Specifically, the invention relates to a salivary gland tissue-specific high-efficiency integrated transgene carrier and its construction method. The invention also relates to a PCR amplification fragment of the mouse parotid gland protein upstream regulatory region and its cloning method . Background technique [0002] Traditional vector construction mainly uses genetic engineering restriction endonuclease, T4 ligase and PCR technology to connect the target fragment to the corresponding vector backbone, and then completes the vector construction after enzyme digestion identification and sequencing verification. According to the needs of the experiment, sometimes multiple fragments need to be ligated, often due to the restriction of the restriction enzyme site, the experimental design becomes more and more difficult, the larger and larger the vector fragments lead to the low efficiency of T4 ligase, ...

AI Technical Summary

Problems solved by technology

Although these two vectors can be used for drug screening, there are no visible markers, and it is difficult to confirm the screening of transgenic cells. They are still traditional vectors, and their integration mode is similar to that of the mouse parotid protein vector constructed by SergueiP.Golovan

However, due to the lack of thorough research on the porcine parotid protein promoter, the current analysis shows that in its upstream 10Kb regulatory region, the regulatory elements are incomplete, and there may be other enhancer regions in other regions, resulting in low expression of the downstream protease gene

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