2005Journal of Sichuan UniversityRequires access

The cDNA Cloning of a Gene Encoding an Unknown Protein from Brassica napus

Fan Zhang

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Abstract

According to the sequence of a 352bp fragment-M17 from the suppression subtractive hybridization library of the mutant line Cr3529 of Brassica napus,primers were designed and a 1000bp upstream fragment was cloned by 5'-RACE,which was joined with the original piece and composed a 1343bp sequence.According to the 3' and 5' ends sequences,a pair of primers were designed and the encoding region sequence of this unknown protein was amplified and cloned.Sequence analyzing showed that it had 80.1% identity with an unknown-protein-encoding gene from Arabidopsis thaliana.Swiss-model was used to predict the structure of the induced protein,and a potential transmembrane helix from site 46 Leu to 66 Phe was identified.GLOBE prediction of globularity showed that this protein possessed a compact globular domain.

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What this paper is about

According to the sequence of a 352bp fragment-M17 from the suppression subtractive hybridization library of the mutant line Cr3529 of Brassica napus,primers were designed and a 1000bp upstream fragment was cloned by 5'-RACE,which was joined with the original piece and composed a 1343bp sequence.According to the 3' and 5' ends sequences,a pair of primers were designed and the encoding region sequence of this unknown protein was amplified and cloned.Sequence analyzing showed that it had 80.1% identity with an unknown-protein-encoding gene from Arabidopsis thaliana.Swiss-model was used to predict the structure of the induced protein,and a potential transmembrane helix from site 46 Leu to 66 Phe was identified.GLOBE prediction of globularity showed that this protein possessed a compact globular domain.

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Available abstract

According to the sequence of a 352bp fragment-M17 from the suppression subtractive hybridization library of the mutant line Cr3529 of Brassica napus,primers were designed and a 1000bp upstream fragment was cloned by 5'-RACE,which was joined with the original piece and composed a 1343bp sequence.According to the 3' and 5' ends sequences,a pair of primers were designed and the encoding region sequence of this unknown protein was amplified and cloned.Sequence analyzing showed that it had 80.1% identity with an unknown-protein-encoding gene from Arabidopsis thaliana.Swiss-model was used to predict the structure of the induced protein,and a potential transmembrane helix from site 46 Leu to 66 Phe was identified.GLOBE prediction of globularity showed that this protein possessed a compact globular domain.

Key concepts: Complementary DNA, Genetics, Gene, Biology, Brassica, Suppression subtractive hybridization, Cloning (programming), Sequence analysis

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