• Asian-Australasian Journal of Animal Sciences
• /
• v.23 no.3
• /
• pp.292-301
• /
• 2010

Follicle-stimulating hormone (FSH) is a pituitary glycoprotein hormone that is encoded by separate alpha- and betasubunit genes. It plays a key role in stimulating and regulating ovarian follicular development and egg production in chicken. FSH signal transduction is mediated by the FSH receptor (FSHR) that exclusively interacts with the beta-subunit of FSH, but characterization of prokaryotic expression of the FSHb gene and its effect on the expression of the FSHR gene in local chickens have received very little attention. In the current study, the cDNA fragment of the FSHb gene from Dagu chicken was amplified using reverse transcription polymerase chain reaction (RT-PCR), and inserted into the pET-28a (+) vector to construct the pET-28a-FSHb plasmid. After expression of the plasmid in E. coli BL21 (DE3) under inducing conditions, the recombination protein, $FSH{\beta}$ subunit, was purified and injected into the experimental hens and the effect on the mRNA expression levels of the FSHR gene was investigated. Sequence comparison showed that the coding region of the FSHb gene in the local chicken shared 99%-100% homology to published nucleotides in chickens; only one synonymous nucleotide substitution was detected in the region. The encoded amino acids were completely identical with the reported sequence, which confirmed that the sequences of the chicken FSHb gene and the peptides of the $FSH{\beta}$ subunit are highly conserved. This may be due to the critical role of the normal function of the FSHb gene in hormonal specificity and regulation of reproduction. The results of gene expression revealed that a recombinant protein with a molecular weight of about 19 kDa was efficiently expressed and it was identified by Western blotting analysis. After administration of the purified $FSH{\beta}$ protein, significantly higher expression levels were demonstrated in uterus, ovary and oviduct samples (p<0.05). These observations suggested that the expressed $FSH{\beta}$ protein possesses biological activity, and has a potential role in regulation of reproductive physiology in chickens.

• Journal of Life Science
• /
• v.21 no.11
• /
• pp.1652-1657
• /
• 2011

The production of blood cells is regulated by more than 20 different growth factors, called hematopoitic growth factors. These factors have been produced in prokaryotic and mammalian systems for their clinical use. Glranulocyte-Colony Stimulating Factor (G-CSF) is an important therapeutic factor for cancer patients as well as patients with congenital conditions. These patients do not have enough neutrophils and have a high risk of infection. Two groups of recombinant G-CSF have been used to specially treat cancer patients after chemotherapy because chemotherapy induces neutropenia, a major side effect of chemotherapy drugs. Here, structural and biological characteristics of G-CSF are presented. In addition, the relationship between chemotherapy and neutropenia, which is a severe reduction of neutrophils in the blood, and clinical application of G-CSF is discussed. Recombinant G-CSFs are grouped in two forms. Non-glycosylated G-CSF, filgrastim, is produced in Escherichia coli and glycosylated G-CSF, lenograstim, is produced in Chinese hamster ovary cells. Differences in structure and biological activity are compared and challenges for biosimilar production are also highlighted.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.42 no.7
• /
• pp.1109-1114
• /
• 2013

Microbial community analysis was performed on Tarak, a traditional Korean fermented milk product, by 16S rDNA cloning and pyrosequencing to obtain basic data for the standardization and systematization of the Tarak manufacturing process. Microbial analysis of the prokaryotic community revealed a slight difference in microbial abundance between Bontarak (n) and Tarak (n+1), but Firmicute was dominant at the phylum level. At the genus level, the Lactobacillus and Leuconostoc genera constituted over 90% of the population in Bontarak, but Lactococcus was the dominant genus in Tarak. Bontarak and Tarak showed further differences at the species level. Leuconostoc citreum was the dominant species in Bontarak, constituting 40% of the population. In eukaryotic community analysis, all samples were composed of Ascomycota at the phylum level. At the genus level, Saccharomyces was dominant in Bontarak (85% of the population), while Issatchenkia was dominant in Tarak (95% of the population). At the species level, Saccharomyces cerevisiae was detected at a relative abundance in Bontarak (82%), and Pichia kudriavzevii was the dominant species in Tarak, with a relative abundance of 95%. Sensory evaluation indicated that Tarak had a better appearance and texture than Bontarak. As sweetness was not significantly different between the two samples just slightly higher in Tarak, this was likely due to a significant decrease in sourness in Tarak. These results suggest that the microbial community used affects the quality of Tarak produced. Thus, a stable microbial community must be maintained for the production of Tarak with consistent quality.