| 摘要: |
| 雨生红球藻(Haematococcus pluvialis)的β-胡萝卜素酮化酶是虾青素生物合成途径中关键酶之一,本研究通过基因枪法将克隆出的β-胡萝卜素酮化酶基因(bkt),转入莱茵 衣藻(Chlamydomonas reinhardtii)的叶绿体中,经过含100mg/L壮观霉素固体培养基筛选得到转化子。通过PCR和基因组酶切Southern杂交分析,证明 bkt基因通过同源重组定点整合到转化子的叶绿体基因组中。进一步通过RT-PCR和RT-PCR Southern杂交分析,表明bkt在衣藻转化子中得到表达。 |
| 关键词: 莱茵衣藻(Chlamydomonas reinhardtii) 叶绿体转化 β-胡萝卜素酮化酶基因(bkt) |
| DOI： |
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| 基金项目:中国科学院知识创新工程资助项目(KZCXZ-YW-209) |
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| Expression of β-carotene ketolase gene from the green alga Haematococcus pluvialis in Chlamydomonas reinhardtii chloroplast |
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| Abstract: |
| β-carotene ketolase from Haematococcus pluvialis is one of the key enzymes of the astaxanthin biosynthesis. Chlamydomonas reinhardtii chloroplast expression vector p64Dbkt contained β-carotene ketolase gene (bkt) under the atpA promoter and rbcL terminator. The vector p64Dbkt was transferred to the chloroplast genome of C. reinhardtii using a micro-particle bombardment. The transgenic alga was selected and maintained in the TAP agar plates containing 100 mg/L spectinomycin. PCR and Southern blot analyses indicated that bkt was integrated into the chloroplast genome of the transformants. RT-PCR and RT-PCR Southern assays showed that bkt gene was expressed in C. reinhardtii transformants. |
| Key words: Chlamydomonas reinhardtii chloroplast transformation β-carotene ketolase gene (bkt) |
