RNA干扰沉默PID1基因肉兔模型的构建

doi:10.11843/j.issn.0366-6964.2014.05.010

Abstract

Abstract:

For the further research of PID1 gene function，transgenic rabbit model was constructed by use of sperm-mediated gene transfer on the basis of the PID1-shRNA expression vector pGPU6/GFP/Neo-PID1-2 preliminarily built and screened.Offsprings were detected by fluorescence detection in vivo，PCR，RT-PCR and Western blot.Individuals from different groups(positive group，negative group and blank group) were selected at the weight of about 3.0 kg and slaughtered to determine their intramuscular fat contents.The results showed that exogenous gene and green fluorescent protein (GFP) gene can be successfully expressed in transgenic offspring rabbits with transgenic positive rate 6.60% (7/106).Compared with the negative and blank groups，the mRNA expression level of the positive rabbits decreased significantly which was detected by RT-PCR (P<0.05).Western blot detection indicated that protein expression level showed a downward trend.Intramuscular fat content also decreased significantly (P<0.05).The results indicated that PID1 gene was closely related to the intramuscular fat deposition.The study further validated the function of PID1 gene in intramuscular fat deposition from the perspective of RNA interference model.What’s more，the study laid the foundation for preparation of high quality PID1 transgenic pigs with high intramuscular fat content.

CLC Number:

XU Zheng-gang， YANG Lun， ZENG Yong-qing， ZHANG Zhe， CHEN Wei， YANG Yun， FANG Guo-feng， WANG Shou-dong. Construction of Transgenic Rabbit Model by RNA Interferencing PID1 Gene[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(5): 750-756.

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Construction of Transgenic Rabbit Model by RNA Interferencing PID1 Gene

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