Comparing and combining distance-based and character-based approaches for barcoding turtles

Molecular barcoding can serve as a powerful tool in wildlife forensics and may prove to be a vital aid in conserving organisms that are threatened by illegal wildlife trade, such as turtles (Order Testudines). We produced cytochrome oxidase subunit one (COI) sequences (650 bp) for 174 turtle species and combined these with publicly available sequences for 50 species to produce a data set representative of the breadth of the order. Variability within the barcode region was assessed, and the utility of both distance-based and character-based methods for species identification was evaluated. For species in which genetic material from more than one individual was available (n = 69), intraspecific divergences were 1.3% on average, although divergences greater than the customary 2% barcode threshold occurred within 15 species. High intraspecific divergences could indicate species with a high degree of internal genetic structure or possibly even cryptic species, although introgression is also probable in some of these taxa. Divergences between species of the same genus were 6.4% on average; however, 49 species were <2% divergent from congeners. Low levels of interspecific divergence could be caused by recent evolutionary radiations coupled with the low rates of mtDNA evolution previously observed in turtles. Complementing distance-based barcoding with character-based methods for identifying diagnostic sets of nucleotides provided better resolution in several cases where distance-based methods failed to distinguish species. An online identification engine was created to provide character-based identifications. This study constitutes the first comprehensive barcoding effort for this seriously threatened order.     

外源质粒pcDNA3s疫苗在小鼠体内组织代谢动力学研究

为分析外源质粒pcDNA3s肌肉注射后在小鼠体内的组织分布及评价整合到宿主基因组上的可能性．通过PCR方法检测质粒pcDNA3s经肌肉注射在各组织中的分布及随时间变化的情况．检测外源质粒在基因组DNA上的整合情况．注射后3h所有的组织均能检测到质粒,至第3周时只有胸腺和生殖器官能检测到质粒的存在,至第6周后各组织均为阴性、各组织基因组DNA经PCR扩增均为阴性．肌注外源质粒后,外源质粒在小鼠细胞内逐渐被清除,外源质粒不会整合到宿主染色体上．