Identifikacija ptičjih vrsta lančanom reakcijom polimerazom i analizom slijeda mitohondrijskoga gena 12S rRNA

Authentic identification and differentiation of avian species is a vital step in conservative, taxonomic, forensic, legal and other ornithological interventions. The present investigation involved the application of molecular biological approach to identify and differentiate avian species i.e. two species of birds, namely black kite (Milvus migrans) and parakeet (Psittacula krameri). The DNA was isolated from blood samples of each species and a part of the mitochondrial 12S rRNA gene was amplified through polymerase chain reaction (PCR). The PCR products were sequenced and aligned using Basic Local Alignment Search Tool (BLAST) of the GenBank (NCBI). Based on the alignment and similarity/divergence, these avian species were accurately identified and differentiated.Autentična identifikacija i razlikovanje ptičjih vrsta od presudnoga su značenja u različitim konzervirajućim, taksonomskim, sudbenim, zakonskim i drugim ornitološkim aktivnostima. Ovo istraživanje bavi se molekularnobiološkim pristupom identifikaciji i razlikovanja dviju ptičjih vrsta: crvenkaste lunje (sokola) (Milvus migrans) i papige (Psittacula krameri). DNA je bila izdvojena iz uzoraka njihove krvi te je dio mitohondrijskoga 12S rRNA bio umnožen lančanom reakcijom polimerazom. Proizvodi PCR-a bili su sekvencirani i analizirani upotrebom Basic Local Alignment Search Tool (BLAST) genske banke GenBank (NCBI). Na osnovi sličnosti odnosno različitosti nalaza identificirane su te dvije pretraživane vrste

Evaluation of PEGylation reaction and purification of monoPEGylated recombinant human granulocyte colony stimulating factor

1049-1053In this study, incubation of methoxy polyethylene glycol propionaldehyde (mPEG-ALD) with recombinant human granulocyte colony stimulating factor (rh-GCSF) in presence of  cyanoborohydride yielded various species of polyethylene glycol (PEG) conjugated GCSF as detected by size exclusion high performance liquid chromatography (SE-HPLC). These reactions were carried out at 1:5 molar ratio of rh-GCSF to PEG at 20-25°C. At 1 h of reaction, mainly monoPEGylated GCSF (62.4%) was formed. At 2 h of reaction, multi PEGylated GCSF (conc. 9%) was detected, where monoPEGylated GCSF constituted the most (73.2%). At 3 h of reaction, monoPEGylated (76.2%) and multi PEGylated GCSF (11.3%) were formed. Further, a process involving ion exchange and gel filtration chromatography were used to obtain pure monoPEGylated GCSF. Purified monoPEGylated GCSF was comparable to standard PEGylated rh-GCSF on NFS-60 cell line, suggesting retained biological activity of monoPEGylated GCSF. Further, the procedure is warranted for purification of other monoPEGylated proteins for therapeutic purpose