初探lncRNA-Tubb4b的蛋白编码能力及其对微管基因的调控

doi:10.11843/j.issn.0366-6964.2022.01.018

Abstract

Abstract: The study aimed to explore whether the long non-coding RNA of Tubb4b (lncRNA-Tubb4b) has protein coding capability and the regulatory effect of lncRNA-Tubb4b on the Tubb4b gene. The ATG, 0/1/2 T tails and Flag sequence were added to lncRNA-Tubb4b to analyze the coding ability of lncRNA-Tubb4b by constructing vector, liposome transfection and Western blot; Then, overexpression vector of lncRNA-Tubb4b was constructed and the siRNA of lncRNA-Tubb4b was synthesized, while the liposome transfection and real-time fluorescent quantitative PCR technology were used to detect the influence of lncRNA-Tubb4b on the Tubb4b gene.The results showed that the Flag fragments with 0, 1, and 2 T-tail, was amplified in this study, were fused with lncRNA-Tubb4b, then were successfully constructed into the pcDNA3.1(-) vector; After transfection of mouse spermatogonia, it was found that lncRNA-Tubb4b with 0, 1, and 2 T-tail had no FLAG protein bands. With pcDNA 3.1(-)-EGFP vector as a control, overexpression of lncRNA-Tubb4b significantly reduced the mRNA expression level of Tubb4b in mouse spermatogonia (P<0.01), while there was fluorescence in control cells. And interfering the expression of lncRNA-Tubb4b significantly increased the mRNA expression level of Tubb4b in mouse spermatogonia (P<0.01). It is indicated that lncRNA-Tubb4b does not encode protein, and overexpression or interference with the expression of lncRNA-Tubb4b will extremely significantly reduce or increase the expression of Tubb4b.

Key words: lncRNA-Tubb4b, tubule, Tubb4b, protein coding, mouse

CLC Number:

S814.1

FENG Meiying, WEI Hengxi, LI Li, ZHANG Shouquan. The Preliminary Research of the Protein Coding Ability of lncRNA-Tubb4b and Its Regulation on Microtubule Genes[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(1): 179-187.

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The Preliminary Research of the Protein Coding Ability of lncRNA-Tubb4b and Its Regulation on Microtubule Genes

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