Recombinant adenovirus ADV-lncRNA Gm44275 as well as construction method and application thereof
A technology of adv-lncrnagm44275 and recombinant adenovirus, which is applied in the direction of recombinant DNA technology, viruses, applications, etc., can solve the problems affecting meat food safety, exogenous hormone residues, and high requirements for precise delivery control, etc., to achieve precise control of female animal delivery , individual differences have little influence, and the effect is stable
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Embodiment 1
[0044] Example 1LNCRNA constructs of overexpression vectors
[0045] Long chain non-coding RNA (Long Noncoding RNA, LNCRNA) refers to a non-encoded RNA having a length greater than 200 nucleotides. The non-coded RNA plays an important role in an apparent genetic regulation, a cell cycle regulation, and cell differentiation, etc., and some LNCRNA has tissue and spatial specificity. The present invention first synthesizes LNCRNAGM44275 (LNCRNA GM36582) by in vitro synthesis, and sequence sequence sequence sequence (sequence sequence sequence sequence sequence sequence) and green fluorescent label sequence (selected) is then added to the LNCRNA GM44275 sequence. See SEQ ID No. 3) to obtain a lncRNA overexpression vector, which sequence seq ID No.4.
Embodiment 2
[0046] Example 2ADV packaging lncRNA overexpression vector
[0047] Adenovirus (ADV) refers to a macromolecule (36 kB) double-strandless DNA virus. The adenoviral genome was transferred to the nucleus through the receptor-mediated endocytosis. The inactivated adenovirus can be integrated as a carrier, injecting the target gene fragment, injecting the human or animal, promoting the expression of the gene.
[0048] Padeno vecotr vector is an adenovirus expression vector, carrier structure mode such as figure 1 Indicated. The above-mentioned LNCRNA overexpression vector is homologously recombinant in vitro to the Padeno vecotr vector, and the LNCRNAGM44275 (LNCRNA GM36582) sequence is transferred to the Padeno Vecotr vector to give GM44275-CBH-GFP adenovirus plasmid.
[0049] The plasmid was linearized by PACI, and the enzyme digestion was transfected into HEK293 cells, and when the cells had a pathological reaction and 50% left and right cells fall off, lysed cells were collected by...
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