Construction of Plant Binary Expression Vector for OsPIP2; 7 with HA-tag and Its Transformation in Rice

doi:10.7668/hbnxb.201750580

Abstract

Abstract: To analyze the function of OsPIP2; 7 under environmental stresses, specific primers were designed to clone OsPIP2; 7 from the complementary DNA of rice variety Zhonghua 11. The nucleotide sequence of 3×HA tag was connected to 3'-end of OsPIP2; 7 with removed terminator by nested PCR amplification, and then the DNA fragments were inserted into pHB vector to construct pHB-OsPIP2; 7-3×HA. The calli of of mature embryo from Zhonghua 11 were used to conduct the genetic transformation by Agrobacterium -mediated transformation method. After screening and transformation on a culture medium containing hygromycin, the positive transformants were obtained. Western-blot results revealed that OsPIP2;7-HA was over-expressed in transgenic plant at protein level when using HA as the first antibody. This study will help the subsequent research of OsPIP2; 7 function and mechanism.

Key words: Rice, Aquaporin, HA tag, Protein expression

CLC Number:

Q785

WANG Xiaojing, SUN Linjing, SUN Yue, ZHANG Rongxue, LI Junling, YAN Shuangyong, SU Jingping. Construction of Plant Binary Expression Vector for OsPIP2; 7 with HA-tag and Its Transformation in Rice[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(5): 59-64. doi: 10.7668/hbnxb.201750580.

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