Expression of bkt and bch genes from Haematococcus pluvialis in transgenic Chlamydomonas  

作　　者：ZHENG KaiJing WANG ChaoGang XIAO Ming CHEN Jun LI JianCheng HU ZhangLi 

机构地区：[1]Shenzhen Key Laboratory of Marine Bioresource & Eco-environmental Science, Shenzhen Engineering Laboratory for Marine Algal Biotechnology, College of Life Science, Shenzhen University

出　　处：《Science China(Life Sciences)》2014年第10期1028-1033,共6页中国科学（生命科学英文版）

基　　金：supported by the National Natural Science Foundation of China(41176106,31470389,31470431);Shenzhen Grant Plan for Science & Technology(CXB201104210005A,JCYJ20120613112512654,JSGG20130411160539208);Guangdong Enterprise Academician Workstation(2011A090700015)

摘　　要：β-carotene ketolase and β-carotene hydroxylase encoded by bkt and bch, respectively, are key enzymes required for astaxanthin biosynthesis in Haematococcu pluvialis 34-1n. Two expression vectors containing cDNA sequences of bkt and bch were constructed and co-transformed into cell-wall-deficient Chlamydomonas reinhardtii CC-849. Transgenic algae were screened on TAP agar plates containing 10 gg mL 1 Zeomycin. PCR-Southern analysis showed that bkt and bch were integrated into the genomes of C. reinhardtii. Transcripts of bkt and bch were further confirmed by RT-PCR-Southern analysis. Compared with the wild type, transgenic algae produced 29.04% and 30.27% more carotenoids and xanthophylls, respectively. Moreover, the transgenic algae could accumulate 34% more astaxanthin than wild type. These results indicate that foreign bkt and bch genes were successfully translated into β-carotene ketolase and β-carotene hydroxylase, which were responsible for catalyzing the biosynthesis of astaxanthin in transgenic algae.β-carotene ketolase andβ-carotene hydroxylase encoded by bkt and bch,respectively,are key enzymes required for astaxanthin biosynthesis in Haematococcu pluvialis 34-1n.Two expression vectors containing cDNA sequences of bkt and bch were constructed and co-transformed into cell-wall-deficient Chlamydomonas reinhardtii CC-849.Transgenic algae were screened on TAP agar plates containing 10μg mL-1 Zeomycin.PCR-Southern analysis showed that bkt and bch were integrated into the genomes of C.reinhardtii.Transcripts of bkt and bch were further confirmed by RT-PCR-Southern analysis.Compared with the wild type,transgenic algae produced 29.04%and 30.27%more carotenoids and xanthophylls,respectively.Moreover,the transgenic algae could accumulate 34%more astaxanthin than wild type.These results indicate that foreign bkt and bch genes were successfully translated intoβ-carotene ketolase andβ-carotene hydroxylase,which were responsible for catalyzing the biosynthesis of astaxanthin in transgenic algae.

关 键 词：CHLAMYDOMONAS β-carotene ketolase β-carotene hydroxylase carotenoids XANTHOPHYLLS ASTAXANTHIN 

分 类 号：Q943.2[生物学—植物学]