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| Cloning and Expression of SRK2F Gene in Chinese Cabbage |
| ZHOU Zhi-guo，WANG Cong-yan |
| （Life Science Department，Langfang Normal college，Langfang 065000，Hebei，China） |
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Abstract Taking non salt and drought tolerant inbred lines of Chinese Cabbage〔Brassica campestris L.spp. pekinensis（Lour）Olsson〕SY-14-06 as material，extracted total RNA from root，and reverse transcribed cDNA．According to SRK2F gene sequence of Brassica rapa，primers were designed and 1 044 bp open reading frame（ORF）was cloned．The SRK2F protein contained 347 aminoacids，with a prediction molecular weight of 39.3 kD and PI of 4.88．We construct the procayotic expressive plasmids pEASY-E1-SRK2F using pEASY-E1 vector．After transformation to Transetta（DE3），the expression of recombinant proteins were detected with SDSPAGE．The structural analysis of SRK2F though Smart-embl showed that it contained silk threonine structural domain，which was located at the position of 4-260 aminoacid residues．The ClustalX2 comparison indicated that the SRK2F had close genetic relationship with Arabidopsis thaliana．Finally，purified this protein by affinity chromatography medium，which was known as Nickel ion metal affinity chromatography medium．Thus，gained the purified fusion protein.
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Received: 05 July 2014
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