Directed Duplication of CaMV35S Promoter Influerues the Expression of the Foreign Genes in Transgenic Plants
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Directed Duplication of CaMV35S Promoter Influerues the Expression of the Foreign Genes in Transgenic Plants
Acta Scientiarum Naturalium Universitatis SunYatseniVol. 34, Issue 2, Pages: 60-67(1995)
作者机构:
1. 中山大学生物工程研究中心
2. 广州市教育学院生物学系
作者简介:
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DOI:
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Published:1995,
Published Online:25 March 1995,
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Yan Xiaolan, Qiu Guohua, Li Baojian. Directed Duplication of CaMV35S Promoter Influerues the Expression of the Foreign Genes in Transgenic Plants. [J]. Acta Scientiarum Naturalium Universitatis SunYatseni 34(2):60-67(1995)
DOI:
Yan Xiaolan, Qiu Guohua, Li Baojian. Directed Duplication of CaMV35S Promoter Influerues the Expression of the Foreign Genes in Transgenic Plants. [J]. Acta Scientiarum Naturalium Universitatis SunYatseni 34(2):60-67(1995)DOI:
Directed Duplication of CaMV35S Promoter Influerues the Expression of the Foreign Genes in Transgenic Plants
The mediated vector pLB38 was constructed by insertion the 800 bp fragment of CaMV35S promoter from the plasmid pBI 121 into the BamH I site between the CaMV 35S pro- moter and GUS gene of the pBI121 and screening the recombinants containing the directed dupli- cate of CaMV35S promoter with Xba I digest.The plasmid pBI121 and pLB38 were introduced into the Agrobacterium tumefaciens containing Ti plasmid pGV 3850 by triparental-mating tech- nique with the aid of helper plasmid pGJ23. The foreign genes in pLB38 and pBI121 were transferred into the tobcco plants using the leaf -disk co-cultivated method and two kinds transgenic plants were obtained. DNA / DNA dot blot and Southern blot with α-32P labelled probe containing 3kb fragment of CaMV 35S-GUS comfirmed that foreign genes were transferred and integrated into the trans- genic tobacco plant genome
The identification of the product of NPT II gene with NPT II dot as- say
GUS gene with GUS fluorescent assay proved that the foreign NPT II and GUS gene were expressed in transgenic plants.And the GUS activity in pLB38-transformed plants in which the GUS gene was controlled by two directed CaMV35S promoters was three-to four-fold higher than that in the pBI121-transformed plants in which the GUS gene is controlled by single CaMV35S promoter.These results suggested that the promoter number has effect on the expres- sion level of genes controlledy by these promoters.