渔业研究 ›› 2023, Vol. 45 ›› Issue (6): 593-602.DOI: 10.14012/j.cnki.fjsc.2023.06.010
收稿日期:
2023-04-14
出版日期:
2023-12-25
发布日期:
2023-12-07
通讯作者:
郑明敏(1991—),女,副教授,硕士研究生导师,博士,研究方向为微藻生物技术。E-mail: 作者简介:
张甜甜(1998—),女,硕士研究生,研究方向为微生物学。E-mail:2803797937@qq.com
基金资助:
ZHANG Tiantian1(), CHEN Bilian1,2, ZHENG Mingmin1,2,*()
Received:
2023-04-14
Online:
2023-12-25
Published:
2023-12-07
摘要:
小球藻是一种可以利用阳光和二氧化碳进行光合生长并积累生物质的单细胞微藻,被广泛用于生产生物燃料和其他高附加值产品,在水产养殖、食品、能源和环境等领域均有重要应用。随着基因工程技术的发展,小球藻分子水平方面的研究和应用逐渐受到关注,对其进行特定的遗传改造也成为了可能。本文通过从转化系统、转化方法、转化影响因素、外源蛋白表达等四个方面进行总结归纳,阐述了小球藻在基因工程改造领域中的应用和仍存在的难题,期望为小球藻的高附加值产品生产提供理论支持和参考。
中图分类号:
张甜甜, 陈必链, 郑明敏. 小球藻的基因工程改造研究进展[J]. 渔业研究, 2023, 45(6): 593-602.
ZHANG Tiantian, CHEN Bilian, ZHENG Mingmin. Advances of genetic engineering modification in Chlorella[J]. Journal of Fisheries Research, 2023, 45(6): 593-602.
小球藻藻种 Chlorella species | 抗性基因 Resistant genes | 抗生素 Antibiotic | 最佳筛选浓度/mg·mL-1 Optimal screening concentration | 参考文献 Reference |
---|---|---|---|---|
普通小球藻C.vulgaris | aadA | 奇霉素 | 0.10~0.20 | [ |
椭圆小球藻C.ellipsoidea | NptⅡ | G418 | 0.03 | [ |
蛋白核小球藻C.pyrenoidosa | aadA | 硫酸链霉素 | 0.10 | [ |
蛋白核小球藻C.pyrenoidosa | 卡那霉素 | 1.00 | [ | |
小球藻Chlorella | Kanr | G418 | 1.00 | [ |
椭圆小球藻C.ellipsoidea | Sh ble | 腐草霉素 | 0.001 | [ |
普通小球藻C.vulgaris | hpt | 潮霉素 | 0.02 | [ |
小球藻Chlorella sp. | hpt | 潮霉素B | 0.05 | [ |
普通小球藻C.vulgaris | hpt | 潮霉素B | 0.30 | [ |
蛋白核小球藻C.pyrenoidosa | NptⅡ | G418 | 0.03 | [ |
普通小球藻C.vulgaris | NptⅡ | G418 | 0.03 | [ |
椭圆小球藻C.ellipsoidea | NptⅡ、NR | G418 | 0.03 | [ |
椭圆小球藻C.ellipsoidea | ble | 博来霉素 | 0.01 | [ |
普通小球藻C.vulgaris | CAT | 氯霉素 | 0.20 | [ |
普通小球藻C.vulgaris | hpt | 潮霉素B | 0.02 | [ |
小球藻Chlorella | neo | G418 | 0.16 | [ |
表1 用于不同种类小球藻的抗生素抗性基因及最佳筛选浓度
Tab.1 The resistant genes and optimal screening concentration of antibiotic for different species of Chlorella
小球藻藻种 Chlorella species | 抗性基因 Resistant genes | 抗生素 Antibiotic | 最佳筛选浓度/mg·mL-1 Optimal screening concentration | 参考文献 Reference |
---|---|---|---|---|
普通小球藻C.vulgaris | aadA | 奇霉素 | 0.10~0.20 | [ |
椭圆小球藻C.ellipsoidea | NptⅡ | G418 | 0.03 | [ |
蛋白核小球藻C.pyrenoidosa | aadA | 硫酸链霉素 | 0.10 | [ |
蛋白核小球藻C.pyrenoidosa | 卡那霉素 | 1.00 | [ | |
小球藻Chlorella | Kanr | G418 | 1.00 | [ |
椭圆小球藻C.ellipsoidea | Sh ble | 腐草霉素 | 0.001 | [ |
普通小球藻C.vulgaris | hpt | 潮霉素 | 0.02 | [ |
小球藻Chlorella sp. | hpt | 潮霉素B | 0.05 | [ |
普通小球藻C.vulgaris | hpt | 潮霉素B | 0.30 | [ |
蛋白核小球藻C.pyrenoidosa | NptⅡ | G418 | 0.03 | [ |
普通小球藻C.vulgaris | NptⅡ | G418 | 0.03 | [ |
椭圆小球藻C.ellipsoidea | NptⅡ、NR | G418 | 0.03 | [ |
椭圆小球藻C.ellipsoidea | ble | 博来霉素 | 0.01 | [ |
普通小球藻C.vulgaris | CAT | 氯霉素 | 0.20 | [ |
普通小球藻C.vulgaris | hpt | 潮霉素B | 0.02 | [ |
小球藻Chlorella | neo | G418 | 0.16 | [ |
小球藻藻种 Chlorella species | 表达载体 Vector | 启动子 Promoter | 转化方法 Transformation method | 转化率或蛋白表达量 Transformation rate or protein expression | 参考文献 Reference |
---|---|---|---|---|---|
普通小球藻 C.vulgaris | pUC改造的 pCMCC | Prrn | 电穿孔 | 人碱性成纤维细胞生长因子表达量0.26~1.42 ng/g藻细胞鲜重 | [ |
普通小球藻 C.vulgaris | pMD18T | P16S、rbcL | 微粒子轰击 | 转化率(212±48)转化子/μg质粒DNA | [ |
索罗金小球藻 C.sorokiniana | pCAMBIA1301 | CaMV35S | 农杆菌介导 | 转化率(220±5)转化子/106细胞 | [ |
普通小球藻 C.vulgaris | pBYR2e | CaMV35S | 农杆菌介导 | 表达量1.14 μg/g鲜重、成纤维细胞生长因子表达量1.61 ng/g鲜重 | [ |
椭圆小球藻 C.ellipsoidea | pGREEN0029、 pBAC818 | e35s | 纳米载体介导 | 转化率(0.95×102±0.07)转化子/μg质粒DNA | [ |
佐夫色绿藻 C.zofingiensis | pBI121 | CaMV35S | 农杆菌介导 | 虾青素质量比 0.564 mg/g | [ |
蛋白核小球藻 C.pyrenoidosa | pBI121 | 微粒子轰击 | 目的蛋白含量高达55.94% | [ | |
小球藻 Chlorella sp. | pMDC85 | 2×CaMV35S | 电穿孔 | 可溶性总蛋白中目的蛋白含量约占0.052%~0.078% | [ |
蛋白核小球藻 C.pyrenoidosa | pGREENII0029 | Ubi | 电穿孔 | 转化率(101±7)转化子/μg质粒DNA | [ |
普通小球藻 C.vulgaris | pBI121 | CaMV35S | PEG介导 | 转化率(356±30)转化子/μg质粒DNA | [ |
椭圆小球藻 C.ellipsoidea | pGREEN0029 | Ubi | 电穿孔 | 转化率750转化子/μg质粒DNA,蛋白表达量11.42 mg/L | [ |
椭圆小球藻 C.ellipsoidea | pSP124 | Ubi-1、rbcS2 | 纤维素酶处理 | 转化率2.25×103转化子/μg质粒DNA | [ |
表2 不同种类小球藻的转化率或蛋白表达量
Tab.2 The transformation rate or expressed proteins in different species of Chlorella
小球藻藻种 Chlorella species | 表达载体 Vector | 启动子 Promoter | 转化方法 Transformation method | 转化率或蛋白表达量 Transformation rate or protein expression | 参考文献 Reference |
---|---|---|---|---|---|
普通小球藻 C.vulgaris | pUC改造的 pCMCC | Prrn | 电穿孔 | 人碱性成纤维细胞生长因子表达量0.26~1.42 ng/g藻细胞鲜重 | [ |
普通小球藻 C.vulgaris | pMD18T | P16S、rbcL | 微粒子轰击 | 转化率(212±48)转化子/μg质粒DNA | [ |
索罗金小球藻 C.sorokiniana | pCAMBIA1301 | CaMV35S | 农杆菌介导 | 转化率(220±5)转化子/106细胞 | [ |
普通小球藻 C.vulgaris | pBYR2e | CaMV35S | 农杆菌介导 | 表达量1.14 μg/g鲜重、成纤维细胞生长因子表达量1.61 ng/g鲜重 | [ |
椭圆小球藻 C.ellipsoidea | pGREEN0029、 pBAC818 | e35s | 纳米载体介导 | 转化率(0.95×102±0.07)转化子/μg质粒DNA | [ |
佐夫色绿藻 C.zofingiensis | pBI121 | CaMV35S | 农杆菌介导 | 虾青素质量比 0.564 mg/g | [ |
蛋白核小球藻 C.pyrenoidosa | pBI121 | 微粒子轰击 | 目的蛋白含量高达55.94% | [ | |
小球藻 Chlorella sp. | pMDC85 | 2×CaMV35S | 电穿孔 | 可溶性总蛋白中目的蛋白含量约占0.052%~0.078% | [ |
蛋白核小球藻 C.pyrenoidosa | pGREENII0029 | Ubi | 电穿孔 | 转化率(101±7)转化子/μg质粒DNA | [ |
普通小球藻 C.vulgaris | pBI121 | CaMV35S | PEG介导 | 转化率(356±30)转化子/μg质粒DNA | [ |
椭圆小球藻 C.ellipsoidea | pGREEN0029 | Ubi | 电穿孔 | 转化率750转化子/μg质粒DNA,蛋白表达量11.42 mg/L | [ |
椭圆小球藻 C.ellipsoidea | pSP124 | Ubi-1、rbcS2 | 纤维素酶处理 | 转化率2.25×103转化子/μg质粒DNA | [ |
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