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2014•Fudan xuebao. Ziran Kexue banRequires access

Cloning and Activity Analysis of a Rice Floral Development Specific Promoter

Wei We

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Abstract

The promoter of a rice EXPANSINfamily gene OsEXPB1 was cloned by Polymerase Chain Reaction(PCR)technique.The 2.7kb sequence before the initiation codon of OsEXPB1 was analyzed online by PLACE software.The results showed that it contains two kinds of cis-elements related to pollen-specific expression.This sequence was fused with GUS reporter gene and transformed into rice.GUS staining results with the positive transgenic plants revealed that GUS signals appear in young developing tissues,but after the plants switch into reproductive growth stage,GUS activity can only be detected in rice florets.There was no signal in all examined vegetative tissues,and the signal only appears in the pollen at the late reproductive development stage.Quantitative reverse transcriptional(qRT)-PCR data confirmed the results of GUS staining.Our results showed that in mature rice plants,OsEXPB1 gene promoter can drive flower-specific gene expression.It may be useful in genetic modification of crop plants.

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What this paper is about

The promoter of a rice EXPANSINfamily gene OsEXPB1 was cloned by Polymerase Chain Reaction(PCR)technique.The 2.7kb sequence before the initiation codon of OsEXPB1 was analyzed online by PLACE software.The results showed that it contains two kinds of cis-elements related to pollen-specific expression.This sequence was fused with GUS reporter gene and transformed into rice.GUS staining results with the positive transgenic plants revealed that GUS signals appear in young developing tissues,but after the plants switch into reproductive growth stage,GUS activity can only be detected in rice florets.There was no signal in all examined vegetative tissues,and the signal only appears in the pollen at the late reproductive development stage.Quantitative reverse transcriptional(qRT)-PCR data confirmed the results of GUS staining.Our results showed that in mature rice plants,OsEXPB1 gene promoter can drive flower-specific gene expression.It may be useful in genetic modification of crop plants.

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Available abstract

The promoter of a rice EXPANSINfamily gene OsEXPB1 was cloned by Polymerase Chain Reaction(PCR)technique.The 2.7kb sequence before the initiation codon of OsEXPB1 was analyzed online by PLACE software.The results showed that it contains two kinds of cis-elements related to pollen-specific expression.This sequence was fused with GUS reporter gene and transformed into rice.GUS staining results with the positive transgenic plants revealed that GUS signals appear in young developing tissues,but after the plants switch into reproductive growth stage,GUS activity can only be detected in rice florets.There was no signal in all examined vegetative tissues,and the signal only appears in the pollen at the late reproductive development stage.Quantitative reverse transcriptional(qRT)-PCR data confirmed the results of GUS staining.Our results showed that in mature rice plants,OsEXPB1 gene promoter can drive flower-specific gene expression.It may be useful in genetic modification of crop plants.

Key concepts: Biology, GUS reporter system, Gene, Cloning (programming), Reporter gene, Pollen, Transgene, Polymerase chain reaction

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