[Relativity betweeen expression level of potato virus X coat protein and the codon usage of wobble codon].
The relationship between the quantities of rare codons and the expression levels of foreign genes is very important in gene engineering. But up to now, there is few experiment on it in multi-cellular eukaryotes, especially in plants. To study the effect of rare codons on the expression level of potato virus X (PVX) coat protein (CP) in transgenic tobacco, some preferred codons in PVX coat protein gene were substituted with synonymous rare codons without changing the encoded amino acid sequence using site-specific mutation. The modified coat protein gene (cpm) and non-modified coat protein gene (cpw) were inserted into binary vector under the control of CaMV35S promoter, and these two plant expression constructs were transferred into tobacco (Nicotiana tabacum cv. Xanthi) genomes via Agrobacterium mediated method and transgenic plants were generated. Western blot and ELISA of these transgenic tobaccos showed that the expression level of modified CP is about one third to one sixth of that of non-modified CP. Northern blot analysis of RNAs from some transformants indicated that the transcriptional levels of cpm are the same as that of cpw, which implicated that the quantity of rare codons in foreign gene was a possible limited factor in foreign gene expression process. Changing the quantity of rare codons maybe an effective method for organism to control gene expression.
Feng
Dejiang
D
Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China. djfeng@genetics.ac.cn
Cai
Huaya
H
Liu
Xiang
X
Xu
Junwang
J
Zhu
Zhen
Z
D000595:Amino Acid Sequence; D001483:Base Sequence; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003062:Codon; D015870:Gene Expression; D008969:Molecular Sequence Data; D015202:Protein Engineering; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism
D036022:Capsid Proteins; D003062:Codon; C058750:coat protein, Potato virus X