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人急性单核细胞白血病相关抗原22RNA干扰序列的筛选及鉴定

Screening and identification of RNA interference sequence for human acute monocytic leukemia related antigen 22
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摘要 目的筛选获得急性单核细胞白血病相关抗原22(MLAA-22)最佳RNA干扰序列,为后期MLAA-22功能研究奠定基础。方法采用反转录聚合酶链反应(RT-PCR)克隆MLAA-22蛋白质编码区序列(CDS),将测序正确的MLAA-22CDS插入pEGFP-N1-3FLAG载体,构建真核pEGFP-N1-3FLAG表达载体,同时构建4个MLAA-22靶向RNA干扰载体,共转染293T细胞后通过细胞荧光强度分析及蛋白印迹法筛选最佳MLAA-22RNA干扰序列。结果成功构建了MIJAA-22真核pEGFP-N1-3FLAG表达载体,同时构建了4个RNA干扰载体,共转染293T细胞后筛选获得KD2为最佳干扰序列。结论KD2是靶向MLAA-22抗原基因的最佳RNA干扰序列。 Objective To screen the optimal RNA interference sequence of acute monocytic leukemia associated antigen 22 (MLAA-22) gene in order to study gene function of it. Methods MLAA-22 coding sequence (CDS) was cloned by reverse transcription polymerase chain reaction (RT-PCR) and the CDS was inserted in to pEGFP-N1-3FLAG vector to construct eukaryotic expression vector of MLAA-22. At the same time, four RNA interference sequences were designed and cloned to the vector. Expression vector and RNA interference vector were co-transfected into 293T cells, and the optimal RNA interference sequence was screened by fluorescence and Western blot analyses. Results MLAA-22 eukaryotic expression vector pEGFP-N1-3FLAG and four RNA interference vectors were successfully constructed. After co-transfected 293T cells, KD2 was selected as the optimal interference sequence of MLAA-22. Conclusion KD2 is an optimal interference sequence for targeting MLAA-22 antigen gene.
作者 古流芳 张鹏宇 曹星梅 张王刚 Gu Liufang, Zhang Pengyu, Cao Xingmei, Zhang Wanggang(Department of Hematology, the Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710004, China)
出处 《白血病.淋巴瘤》 CAS 2018年第3期149-153,共5页 Journal of Leukemia & Lymphoma
基金 国家自然科学基金(81100373)
关键词 白血病 单核细胞 急性 RNA干扰 急性单核细胞白血病相关抗原22 Leukemia, monocytic, acute RNA interference Acute monocytic leukemia associated antigen 22
作者简介 通信作者:张王刚,Email:61176828@qq.com
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