牛lnc RNA SNHG12过表达载体构建

Construction of Lnc RNA SNHG12 overexpression vector

Objective To construct lncRNA SNHG12 overexpression recombinant vector, and to lay the experimental foundation for further exploring the interaction between lncRNA SNHG12, miRNA 491 and CART gene. Methods national center for biotechnology information (NCBI) Gene Database (No. : NC_037329.1) lncRNA SNHG12 sequence was obtained and amplified by polymerase chain reaction (PCR). After double digestion, pcDNA3.1-EGFP vector was ligated to obtain recombinant plasmid PCDNA3.1-EGFP-SnHG12. The pcDNA3.1-EGFP-SNHG12, miRNA 491 and CART plasmids were transfected into 293T cells, and the expression level of lncRNA SNHG12 was detected by quantitative real-time PCR(qRT-PCR). The results showed that the sequence of pcDNA3.1-EGFP-SNHG12 vector was correct. The green fluorescence amount of 293T cells transfected was about 50%, and the intensity was moderate, indicating that the transfection effect was good. qRT-PCR results showed that the expression level of lncRNA SNHG12 in NC group was significantly higher than that in miRNA 491 group in 293T cells. Conclusion pcDNA3.1-EGFP-SNHG12 vector was successfully constructed, and lncRNA SNHG12 had interaction with miRNA491. This study will create experimental conditions for further exploring the network relationship among lncRNA SNHG12, miRNA 491 and CART genes.