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| Cloning，sequence analysis，and prokaryotic expression of Chlamydomonas reinhardtii gene CrPGP3 and CrFAT1 |
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| DOI：10.16768/j.issn.1004-874X.2016.08.027 |
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| Abstract: |
| In order to express the CrPGP3 and CrFAT1 genes of Chlamydomonas reinhardtii in prokaryotic cell，we cloned and analyzed these two sequences. After the total RNA in C. reinhardtii CC124 was extracted，the revers transcription were performed. The full length cDNA obtained by PCR was inserted into the prokaryotic expression vector PGEX-6p-1 and transformed into Escherichia coli BL21（DE3）. These positive clones were called CrPGP3-PGEX-6p-1 and CrFAT1-PGEX-6p-1. The target proteins were expressed after adding 1.0 mmol/L IPTG. The results showed that the CrPGP3 and CrFAT1 gene had 786 bp and 1 188 bp coding 261 and 395 amino acids，respectively. Sequence analysis indicated that CrPGP3 was a member of super family“ CDP-alcohol phosphatidyltransferase”，and the CrFAT1 belonged to a member of“ hot dog” super family. The SDS-PAGE displayed that the expressed proteins were consistent with the anticipated size. The recombinant plasmids expressed the target proteins in E. coli BL21. |
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