| 杜氏盐藻外源基因稳定表达系统的构建(英文) |
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| 引用本文： | 耿德贵,韩燕,王义琴,王鹏,张利明,李文彬,孙勇如.杜氏盐藻外源基因稳定表达系统的构建(英文)[J].Acta Botanica Sinica,2004,46(3):342-346. |
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| 作者姓名： | 耿德贵 韩燕 王义琴 王鹏 张利明 李文彬 孙勇如 |
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| 作者单位： | 中国科学院遗传与发育生物学研究所 北京100101
(耿德贵，王义琴，王鹏，张利明，李文彬)，江苏省徐州生物工程学校 徐州221006
(韩燕)，中国科学院遗传与发育生物学研究所 北京100101(孙勇如) |
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| 摘 要： | A stable transformation system for the expression of foreign genes in the unicellular greenmarine alga (Dunaliella salina Teod.) was established. Using electroporation, the alga was transformed witha plasmid containing the hepatitis B surface antigen (HBsAg） gene and the chloramphenicol acetyltransferase(CAT) gene as a selectable gene. PCR and Southern blotting analysis indicated that the HBsAEgene wasintegrated into the D. salina genome. Northern dotting analysis showed that the HBsAg gene was expressedat the mRNA level. The stable expression of HBsAg protein in transformants was confirmed by HBsAgenzyme-linked immunosorbent assay (HBsAg EUSA) and Western blotting analysis. Also, PCR and Southernblotting analyses showed that the CA Tgene was integrated into the D, salina genome, and CAT EUSAindicated that CAT protein was stably expressed in the cells. The introduced HBsAg DNA and HBsAgprotein expression were stably maintained for at least 60 generations in media devoid of chloramphenicol.This is the first report of the stable expression of foreign genes in D. salina.
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| 关 键 词： | 杜氏盐藻 外源基因 氯霉素乙酰转移酶 乙肝病毒表面抗原 |
Construction of a System for the Stable Expression of Foreign Genes in Dunaliella salina |
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| Authors: | GENGDe-Gui HANYan WANGYi-Qin WANGPeng ZHANGLi-Ming LIWen-Bin SUNYong-Ru |
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| Institution: | [1]InstituteofGeneticsandDevelopmentalBiology,TheChineseAcademyofSciences,Beijing100101,China [2]XuzhouBioengineeringSchoolofJiangsuProvince,Xuzhou221006,China |
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| Abstract: | A stable transformation system for the expression of foreign genes in the unicellular greenmarine alga (Dunaliella salina Teod.) was established. Using electroporation, the alga was transformed witha plasmid containing the hepatitis B surface antigen (HBsAg) gene and the chloramphenicol acetyltransferase(CAT) gene as a selectable gene. PCR and Southern blotting analysis indicated that the HBsAg gene wasintegrated into the D. salina genome. Northern dotting analysis showed that the HBsAg gene was expressedat the mRNA level. The stable expression of HBsAg protein in transformants was confirmed by HBsAgenzyme-linked immunosorbent assay (HBsAg ELISA) and Western blotting analysis. Also, PCR and Southernblotting analyses showed that the CAT gene was integrated into the D. salina genome, and CAT ELISAindicated that CAT protein was stably expressed in the cells. The introduced HBsAg DNA and HBsAgprotein expression were stably maintained for at least 60 generations in media devoid of chloramphenicol.This is the first report of the stable expression of foreign genes in D. salina . |
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| Keywords: | Dunaliella salina hepatitis B surface antigen chloramphenicol acetyltransferase |
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