布鲁氏菌侵染对类泛素SUMO-1表达的影响及类泛素SUMO-1慢病毒表达载体的构建

doi:10.16431/j.cnki.1671-7236.2016.06.004

Abstract

Abstract: To investigate the changes of SUMO-1 (small ubiquitin-related modifier) gene in RAW264.7 cells infected by Brucella,a lentiviral vector for the over expression of mouse SUMO-1 was constructed.RAW264.7 cells was infected by Brucella 16M or M5-90,then quantitative Real-time PCR and Western blotting were used to detect SUMO-1 in RAW264.7 cells.SUMO-1 gene fragment was inserted into the lentiviral vector pLEX-MCS in the use of recombinant,resulting into a recombinant lentivirus pLEX-SUMO-1.Recombinant lentiviral vector was transfected into 293T cells after sequencing correctly,RAW264.7 was infected by supernatant.Quantitative Real-time PCR and Western blotting were used to quantify the transcription of SUMO-1 gene.The results showed that RAW264.7 cells was infected by Brucella 16M or M5-90,SUMO-1 showed a reduced trend in 12 h,after 12 h,SUMO-1 began to rise and became higher than normal levels,and the difference was extremely significant (P < 0.01).The sequencing results showed that mouse SUMO-1 gene lentiviral vector was constructed successfully.The mRNA expression of SUMO-1 gene increased.The results indicated that the expression of SUMO-1 changed after Brucella infection.Over-expression vector targeting SUMO-1 gene was constructed successfully,an experimental basis on the SUMO-1 functional study was provided.

Key words: Brucella; small ubiquitin-related modifier SUMO-1; lentivirus vector; murine macrophages

CLC Number:

LI Qi-feng, DING Jin-hua, DOU Wen-jing, ZHANG Hui, CHEN Chuang-fu. Effect on Expression of SUMO-1 in Cells Infected by Brucella and Construction of Lentivirus Vector that Over-expressing SUMO-1[J]. , 2016, 43(6): 1422-1429.

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Effect on Expression of SUMO-1 in Cells Infected by Brucella and Construction of Lentivirus Vector that Over-expressing SUMO-1

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