The effects of additions/deletions in glycosylated residues of recombinant human EPO (rhEPO) produced in CHO-K1 on their secretion were examined. hEPO cDNA was amplified from human liver mRNA and cloned into the pCR2.1 TOPO. Using overlapping-extension site-directed mutagenesis method, glycosylation sites at 24th, 38th, 83rd, and 126th were respectively or accumulatively removed by substituting its asparagine (or serine) with glutamine. To add novel glycosylation sites, 69 and 105th leucine was mutated to asparagine. Mutant and wild type rhEPO constructs were cloned into the pcDNA3 expression vector with CMV promoter and transfected into CHO cell line, CHO-K1, to produce mutant rhEPO mutant rhEPO proteins. Enzyme-linked immunosorbant assay (ELISA) and Western analysis with monoclonal anti-EPO antibody were performed using supernatants of the cultures showing transient and stable expressions respectively. Addition of novel glycosylation reduced rhEPO secretion dramatically while deletion mutants had little effect except some double deletion mutants (△24/83 and △38/83) and triple mutant (△24/38/83). This fact suggests that not single but combination of changes in glycosyl groups affect secretion of rhEPO in cell culture, possibly via changes in their conformations.

수출 볏짚 제품내에 들어있는 월도 이화명충을 방제하기 위하여 Phostoxin 훈증제를 기온상태에서 들이 프라스틱 chamber를 사용하여 농도 및 처리시간을 달리하여 시험한 결과 다음과 같은 결과를 얻었다. 1. 월동유충은 처리 48시간, 옹은 24시간, 성충은 8시간에서 의 살충율 얻었다. 2. 약제의 사용농도 보다는 처리시간의 장단에 따라서 살충효과의 차이가 있었다.