梅花类黄酮3′-羟化酶基因片段基于基因组DNA的简并PCR法克隆

用该研究设计的“预先去杂-SDS法”从梅花嫩叶提取到高质量的基因组DNA。根据11条已公开发表的并提交到GenBank的类黄酮3＇-羟化酶基因cDNA的假定氨基酸序列的保守区设计2个正向简并引物和3个反向简并引物组成6对引物,仅有1对引物能以PCR法同时从梅花‘南京红须’、‘南京红’和‘粉皮宫粉’的基因组DNA扩增到一个469bp的核苷酸片段,这3个片段在总体上有99．72％的一致性,与11条类黄酮3＇-羟化酶基因cDNA的相应区域有65．57％的一致性。同时,“GGEK”并非类黄酮3＇-羟化酶的特征性模体。这是首次从木本植物的基因组DNA克隆到类黄酮3＇-羟化酶基因片段。该研究结果可为梅花类黄酮3＇-羟化酶基因全长的克隆奠定基础。     

Acacetin antagonized lipotoxicity in pancreatic β-cells via ameliorating oxidative stress and endoplasmic reticulum stress

Molecular Biology Reports - During the pathogenesis and progression of diabetes, lipotoxicity is a major threat to the function and survival of pancreatic β-cells. To battle against the...     

CXCR4/SDF-1 pathway is crucial for TLR9 agonist enhanced metastasis of human lung cancer cell

Accumulating data suggested that CXCR4/SDF-1 pathway may play an important role in the metastasis of tumor. We previously demonstrated that CpG ODN could enhance the metastasis of human lung cancer cell via TLR9. Here we further evaluated the possible role of CXCR4/SDF-1 pathway in the enhanced metastasis of human lung cancer 95D cells induced by CpG ODN. Our data showed down-regulation of CXCR4 expression using siRNA against CXCR4 could significantly reduce the enhanced metastasis of 95D cells induced by CpG ODN both in vitro and in vivo. These results suggested that TLR9 agonist might promote the metastasis of human lung cancer cells via CXCR4/SDF-1 pathway.     

Chromosome banding and FISH with rDNA probe in the diploid and tetraploid loach Misgurnus anguillicaudatus

The chromosomes of the diploid and tetraploid loach Misgurnus anguillicaudatus were analyzed by staining with Ag, chromomycin A₃ (CMA₃)/distamycin A (DA), and DA/4′,6-diamidino-2-phenylindole (DAPI), and using fluorescence in situ hybridization (FISH) with 5.8S + 28S rDNA as a probe. Nucleolus organizer regions (NORs) were mapped to the telomeric region of the short arms of the largest (first) metacentric chromosome pair in the diploid loach with 2n = 50 and the homologous quartet in the tetraploid loach with 4n = 100. The NORs were positive at the same region of the first metacentric chromosome for Ag and CMA₃/DA stainings, but negative for DA/DAPI staining. Four signals at the homologs within the same quartet suggest the duplication of the entire genome from diploid to tetraploid status. However, a size difference was detected between the rDNA signals by FISH and CMA₃ banding.     

Evaluation of antimicrobial activity of certain Chinese plants used in folkloric medicine

Six selected plants, belonging to 3 families from Nanjing of China, were extracted with the solvent 95% (v/v) ethanol to yield 11 extracts. The extracts were evaluated for their effects on the growth of eight clinical bacteria, two fungi and one yeast using a modified agar diffusion method. The results showed that the majority of the extracts investigated showed greater activities against the Gram-positive bacteria than against the Gram-negative bacteria, the fungi and the yeast. The strongest antimicrobial activity was exhibited by the stem extracts of Mahonia fortunei against multiresistant Staphylococcus aureus strains, followed by the stem extracts of Mahonia bealei, while Bacillus thuringiensis was the most sensitive to all extracts.     

The Bryopsis hypnoides plastid genome: multimeric forms and complete nucleotide sequence

Background

Bryopsis hypnoides Lamouroux is a siphonous green alga, and its extruded protoplasm can aggregate spontaneously in seawater and develop into mature individuals. The chloroplast of B. hypnoides is the biggest organelle in the cell and shows strong autonomy. To better understand this organelle, we sequenced and analyzed the chloroplast genome of this green alga.

Principal Findings

A total of 111 functional genes, including 69 potential protein-coding genes, 5 ribosomal RNA genes, and 37 tRNA genes were identified. The genome size (153,429 bp), arrangement, and inverted-repeat (IR)-lacking structure of the B. hypnoides chloroplast DNA (cpDNA) closely resembles that of Chlorella vulgaris. Furthermore, our cytogenomic investigations using pulsed-field gel electrophoresis (PFGE) and southern blotting methods showed that the B. hypnoides cpDNA had multimeric forms, including monomer, dimer, trimer, tetramer, and even higher multimers, which is similar to the higher order organization observed previously for higher plant cpDNA. The relative amounts of the four multimeric cpDNA forms were estimated to be about 1, 1/2, 1/4, and 1/8 based on molecular hybridization analysis. Phylogenetic analyses based on a concatenated alignment of chloroplast protein sequences suggested that B. hypnoides is sister to all Chlorophyceae and this placement received moderate support.

Conclusion

All of the results suggest that the autonomy of the chloroplasts of B. hypnoides has little to do with the size and gene content of the cpDNA, and the IR-lacking structure of the chloroplasts indirectly demonstrated that the multimeric molecules might result from the random cleavage and fusion of replication intermediates instead of recombinational events.