| Autor: |
Zengwen Huang, WuReliHazi Hazihan, Bodai, Baheti, Rizabek, Kadyken, Ulzhan, Nuralieva, Karlygash, Omarova, Juan Zhang, Yaling Gu |
| Zdroj: |
Pakistan Journal of Zoology; Dec2020, Vol. 52 Issue 6, p2353-2361, 9p |
| Abstrakt: |
The Ye Mule Aries (YM) Sheep is a valuable local sheep breed from Xinjiang, China. This breed is well adapted to harsh environmental conditions and displays strong disease resistance, fast growth, and high cold tolerance. The effect of small interfering RNA (siRNA) on the expression of follistatin inhibin alpha gene (INHa) in YM sheep was analyzed, and the INHa gene was amplified from total RNA of the ovary tissue of YM sheep and was successfully cloned. The eukaryotic siRNA expression vector was constructed with the cloned gene as the target and validated at the cellular level. Three RNA interference fragments at different sites were designed based on obtained cDNA of the target gene and the siRNA expression vector pGenesil 10-3p-siRNA was constructed. The siRNA expression vector was digested, identified and sequenced, and then transfected into YM sheep granulosa cells. The sequence of YM sheep INHa gene was obtained and uploaded on the NCBI database (accession number KP-113678.1). The INHa gene from YM sheep is 1109 bp long and translates to approximately 40 KDa protein. Q-PCR analysis showed that the plasmid pGenesil 10-3p-siRNA could interfere with the expression of INHa in granulosa cells to an efficiency of 83%, which was also confirmed through western Blot assay. This study successfully constructed the eukaryotic expression plasmid pGenesil 10-3p-siRNA, and confirmed that the plasmid interfered significantly with the expression of INHa gene in YM sheep. Our current findings can serve as a model for studying the effect of INHa on the breeding performance of YM sheep, providing a novel strategy to improve their reproductive capacities. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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