猪杀菌/通透性增加蛋白基因siRNA载体构建及干扰效果评价

doi:10.11843/j.issn.0366-6964.2015.03.020

Abstract

Abstract:

This study was conducted to construct and select the efficient siRNA interference vector for porcine BPI gene，which provided the foundation for studying the function and mechanism of porcine BPI gene at the cellular level.Referring to the whole coding sequence of porcine BPI gene (GenBank accession number：EF436278)，porcine BPI specific hairpin siRNA fragments were designed and inserted into pcDNA 6.2-GW/EmGFPmiR vector.Four pairs of siRNA expression vectors (RB1，RB2，RB3，RB4) and one pair of negative control (NC) were constructed by PCR and sequencing verification.Then the above vectors were transfected into IPEC-J2 intestinal epithelial cell and further the interference effect was detected.The results showed that 4 BPI-specific siRNA vectors all reduced the BPI gene mRNA expression (P<0.05).Meanwhile the interference effect of RB4 vector was the best，whose efficiency reached 69%.This study successfully screened out the efficient siRNA vector which could exclusively interfere porcine BPI gene expression，which provided experimental basis for further studying the BPI gene function and mechanism for the resistance to gram-negative bacteria infection in porcine intestinal tract at the cellular level.

CLC Number:

S828.2

WU Zheng-chang，YIN Xue-mei，XIA Ri-wei，SUN Shou-yong，ZHU Guo-qiang，WU Sheng-long，BAO Wen-bin. Construction and Evaluation of Porcine Bactericidal/Permeability-increasing Protein Gene (BPI) siRNA Expression Vector[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(3): 491-496.

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Construction and Evaluation of Porcine Bactericidal/Permeability-increasing Protein Gene (BPI) siRNA Expression Vector

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