Cloning and bioinformatic analysis of chalcone isomerase in Scutellaria baicalensis / 中草药

Objective: To clone chalcone isomerase (sbCHI) gene from the callus of Scutellaria baicalensis and to analyze its bioinformatics. Methods: RNA was obtained from scutellariae callus, cDNA was reversely transcribed, specific primers were designed, and then CHI was cloned. The protein characteristics was analyzed using bioinformatics and the phylogenetic tree of CHI was constructed using MEGA5.1. Results: The 648 bp sbCHI (accession number: KP064512) sequence was obtained, which has a complete open reading frame (ORF), encoding an unstable protein with 215 amino acids. The sbCHI encoding protein has isoelectric point (pI) of 5.09 and a calculated molecular weight about 22 980, without transmembrane regions and signal peptide has a conserved domain of chalcone-flavanone isomerase family. In the secondary structure, the percentage of alpha helix, β-extended, and random coil were 37.21%, 23.25%, and 39.54%, respectively. The homologous analysis indicates the nucleotide sequence 99.69% similarity and the amino acid sequence 99.07% similarity with S. baicalensis (ADQ13184.1), only in 31 and 160 were different. Conclusion: The sbCHI in scutellariae callus is successfully cloned, which provides the foundation for further characterization sbCHI functionality and the synthetic biology research of baicalin.

Cloning and bioinformatic analysis and expression analysis of beta-glucuronidase in Scutellaria baicalensis / 中国中药杂志

The β-Glucuronidase gene (sbGUS) cDNA firstly from Scutellari abaicalensis leaf was cloned by RT-PCR, with GenBank accession number KR364726. The full length cDNA of sbGUS was 1 584 bp with an open reading frame (ORF), encoding an unstable protein with 527 amino acids. The bioinformatic analysis showed that the sbGUS encoding protein had isoelectric point (pI) of 5.55 and a calculated molecular weight about 58.724 8 kDa, with a transmembrane regions and signal peptide, had conserved domains of glycoside hydrolase super family and unintegrated trans-glycosidase catalytic structure. In the secondary structure, the percentage of alpha helix, extended strand, β-extended and random coil were 25.62%, 28.84%, 13.28% and 32.26%, respectively. The homologous analysis indicated the nucleotide sequence 98.93% similarity and the amino acid sequence 98.29% similarity with S. baicalensis (BAA97804.1), in the nine positions were different. The expression level of sGUS was the highest in root based on a real-time PCR analysis, followed by flower and stem, and the lowest was in stem. The results provide a foundation for exploring the molecular function of sbGUS involved in baicalcin biosynthesis based on synthetic biology approach in S. baicalensis plants.

Significance of expression of extracellular matrix metalloproteinase inducer in pancreatic cancer / 中华消化杂志

Objective To investigate the expression of extracellular matrix metalloproteinase inducer (EMMPRIN) in pancreatic cancer and its relationship with P-glycoprotein (P-gp) and vascular endothelial growth factor (VEGF),and to study its correlation with invasion,metastasis and drug re- sistance.Methods The expressions of EMMPRIN,P-gp and VEGF in 34 cases of pancreatic cancer were detected by immunohistochemistry.The correlation analysis and Chi squared test were been used by SPSS 10.0.Results The positive rates of EMMPRIN,P-gp and VEGF in pancreatic cancer were 61.8%,55.9% and 64.7% respectively.EMMPRIN,P-gp and VEGF were associated with lymph node metastasis (P＜0.05),but not associated with sex,age,size of tumor and degree of differentia- tion (P＞0.05).There were close relationship between EMMPR1N and P-gp,VEGF respectively (r= 0.398,r=0.432,P＜0.05).Conclusions EMMPRIN,P-gp and VEGF were higher expressed in pancreatic cancer.EMMPRIN express was correlated with both P-gp and VEGF.It has indicated that during the progression of pancreatic cancer,there are close relationship between invasion,metastasis and drug resistance,and EMMPRIN may play key role in this process.