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rotunda through Real-Time quantitative PCR Gene expression of CHS gene in different tissues of B

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Figure 1-1 Research flowchart

Verification of B. rotunda CHS variants with transcriptome analysis Docking of CHS protein with panduratin A

Prediction of secondary and 3D structure of CHS protein

Verification of CHS gene with available sequences on GenBank through BLAST and deposit the B. rotunda CHS gene Study the possibility of variants of CHS gene through sequence analysis

Amplification of complete sequence of CHS gene through Rapid Amplification cDNA Ends

Quantitativ gene expression of CHS gene in different tissues of B. rotunda through Real-Time quantitative PCR Gene expression of CHS gene in different tissues of B. rotunda through reverse transcrition PCR using gene specific primers

Establishment of an optimized RNA extraction method in B. rotunda which suits for gene expression analysis Characterization of core fragment of B. rotunda CHS gene and comparsion with other available CHSs gene on GenBank

Verification of core fragment of B. rotunda CHS gene through BLAST analysis Amplification of core fragment of B. rotunda CHS gene through nested PCR

Primer design through alignment of CHS gene from closely related plants to B. rotunda through manual and online softwares

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