ABSTRACT
<p><b>AIM</b>To investigate the expression of recombinant human phosphodiesterase 3A (HPDE3A) using baculovirus expression system in Tn cell line.</p><p><b>METHODS</b>The HPDE3A cDNA was recombined with baculovirus, and then the recombinant was transfected into Tn cell line. The expression of HPDE3A in Tn cell line was detected and identified by the RT-PCR, SDS-PAGE and Western blotting.</p><p><b>RESULTS</b>The recombinant HPDE3A protein was stably expressed in Tn cell line and detected by the distinct morphological changes of Tn cell, RT-PCR, SDS-PAGE and Western blotting using polyclonal antibody. The M(w) of the recombinant protein was about 120 kD.</p><p><b>CONCLUSION</b>Recombinant HPDE3A can be expressed in Tn cell line using the baculovirus expression system, and thus provided the basic material for studying its bioactivity and application in screening for HPDE3A inhibitor.</p>
Subject(s)
Animals , 3',5'-Cyclic-AMP Phosphodiesterases , Genetics , Metabolism , Baculoviridae , Genetics , Cell Line , Cyclic Nucleotide Phosphodiesterases, Type 3 , Electrophoresis, Polyacrylamide Gel , Moths , Cell Biology , Metabolism , RNA, Messenger , Genetics , Metabolism , Recombinant Proteins , Genetics , Metabolism , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To study the significance of the change of protamine content in the management of male infertility.</p><p><b>METHODS</b>Sperm nuclear proteins extracted from 199 infertile patients were analyzed by polyacrylamide gel-electrophoresis and scanning microdensitometry.</p><p><b>RESULTS</b>Forty-two (21%) cases of the total number had normal sperm nuclear proteins and 157 (79%) had aberrant ones, which mainly presented the interruption of HPRR and abnormality of P2 protamine. Thirty patients were selected at random from 157 abnormal cases for clinical treatment. After the treatment, the sperm nuclear proteins were extracted and analyzed and the results demonstrated that 11 cases (36.6%) improved markedly, 5 (16.6%) restored to normal and the other 16 (46.6%) remained unchanged.</p><p><b>CONCLUSION</b>There is a reduced level or selective absence or even complete selective absence of protamines in infertile patients. Protamines may act as a parameter for evaluating the treatment effect of infertile males and protamine content can be influenced by a certain or several factors.</p>
Subject(s)
Adult , Humans , Male , Electrophoresis, Polyacrylamide Gel , Infertility, Male , Metabolism , Oligospermia , Metabolism , Protamines , Spermatozoa , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression of rat protamine (RP) gene in MEL cells and the effect on cell growth.</p><p><b>METHODS</b>Eukaryotic expression plasmid pCR-3.1-RP was constructed and transfected into MEL cells. The changes of cell growth rate, mitotic index and colony-forming rate in semi-solid medium were investigated.</p><p><b>RESULTS</b>Transfected MEL cells showed lower growth rate, mitotic index and colony-forming rate. Volumes of cells were reduced and reduction of RNA transcription was observed.</p><p><b>CONCLUSION</b>These results suggest that expression of RP in MEL cell may inhibit the cell growth and proliferation.</p>