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| The Cloning, Tissue Expression and Over-expression Study of lncGHR1 Gene in Cong Jiang Xiang Pig (Sus scrofa) |
| CUI Xiao-Zhi1,2, WU Qiao-Qun1,2, WU Xiao-Min1,2, ZHU Xiao-Feng1,2, CHEN Ying-Lian3, XIE Ling-Ling4, ZHAO Jia-Fu1,2, CHEN Wei1,2, XU Hou-Qiang1,2,3,* |
1 Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountains Region of Ministry of Education, Guizhou University/Key Laboratory of Animal Genetics, Breeding and Reproduce in Guizhou Province, Guiyang 550025, China;
2 College of Animal Sciences, Guizhou University, Guiyang 550025, China;
3 School of Medicine, Guizhou University, Guiyang 550025, China;
4 Livestock and Poultry Germplasm Testing Center of Guizhou Province, Guiyang 550025, China |
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Abstract Growth hormone receptor (GHR) plays an important role in animal growth and development. In order to study the expression of long noncoding RNAs (lncRNAs) related to GHR gene in Cong Jiang Xiang pig (Sus scrofa), this paper selected Guizhou Cong Jiang Xiang pig and Yorkshire pig as the research objects, used the growth and development related miR-146b-3p as correlation point, screened out the GHR gene related lncRNAs by SWchen 2.3 program, and verified the lncRNAs by cloning and sequencing. The candidate lncRNA NONSUST017828.1 (GenBank No. TCONS_00020564, named lncGHR1) was used as the further research object. Bioinformatics and tissue expression spectrum analysis were performed, the lncGHR1-CMV over-expression vector was constructed and transfected to human embryonic kidney cells (HEK-293T) and primary myocytes of Cong Jiang Xiang pig, effects of lncGHR1 on the expression of Yes-associated protein (YAP) and transcription activator (TAZ) were detected by qRT-PCR. The results showed that the minimum free energy of the secondary structure of lncGHR1 was -131.20 kcal/mol and contained 3 ORF. The results of local comparison showed that lncGHR1 was the most similar with GHR 3'UTR of pig. The inhibition coefficient of lncGHR1 binding to miRNA was 4.96%, and the score of miR-146-3p with lncGHR1 was the highest. The expression of lncGHR1 was the highest in the spleen and lung of Cong Jiang Xiang pig, which was significantly higher than that in other tissues (P<0.01), the lowest in the heart; the highest expression of Yorkshire was found in liver, followed by lung; the expression of lncGHR1 in heart, spleen, lung, kidney, large intestine, small intestine and longissimus dorsi muscle of Cong Jiang Xiang pig was significantly higher than that of Yorkshire pig (P<0.01), but the expression of lncGHR1 in liver was significantly lower (P<0.01). Over-expression of lncGHR1 in 293T cells and primary myocytes of Cong Jiang Xiang pig significantly increased the expression of TAZ gene (P<0.05), but the expression of YAP gene was not significantly different (P>0.05). These results indicated that lncGHR1 might play a key role in the process of cell proliferation and growth. The above results indicated that lncGHR1 played an important role in the growth and development, and the was significant for the analysis of the genetic mechanism of Cong Jiang Xiang pigs' slow growth and short stature as well as the genetic improvement breeding of pigs.
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Received: 02 April 2020
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Corresponding Authors:
*gzdxxhq@163.com
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