{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,8]],"date-time":"2026-01-08T00:11:29Z","timestamp":1767831089063,"version":"3.49.0"},"reference-count":23,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2002,12,24]],"date-time":"2002-12-24T00:00:00Z","timestamp":1040688000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecular Reproduction Devel"],"published-print":{"date-parts":[[2003,2]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The brushtail possum (<jats:italic>Trichosurus vulpecula<\/jats:italic>) zona pellucida (ZP) is composed of three major glycoproteins, designated ZP1, ZP2, and ZP3 based on their size and homology with eutherian ZP proteins. These proteins are candidate antigens for the development of an immunocontraceptive vaccine to control the fertility of the brushtail possum in New Zealand, where it is an introduced pest. In order to further their immunological and functional characterization, recombinant possum ZP proteins were produced in <jats:italic>Escherichia coli<\/jats:italic> (<jats:italic>E. coli<\/jats:italic>) strain JM109, M15, SG13009, or BL21 codon plus<jats:italic>.<\/jats:italic> Each of the proteins produced possessed a N\u2010terminal six histidine tag (His)<jats:sub>6<\/jats:sub> to facilitate purification and consisted of amino acid (aa) residues 18\u2013471 of possum ZP1, aa residues 40\u2013311 of ZP2 (ZP2\u2010N), aa residues 305\u2013634 of ZP2 (ZP2\u2010C), and aa residues 23\u2013342 of ZP3. Immunoblot using anti\u2010RGS(His)<jats:sub>4<\/jats:sub> antibodies and polyclonal rabbit anti\u2010porcine ZP antibodies detected major bands at 54 kDa for ZP1, 32 kDa for ZP2\u2010N, 39 kDa for ZP2\u2010C, and 40 kDa for ZP3. Immunization of male and female rabbits with ZP2\u2010N, ZP2\u2010C, and ZP3 purified on Ni\u2010NTA resin under denaturing conditions generated antibodies reactive with recombinant ZP proteins on Western blot and with native ZP proteins in possum ovarian sections using immunofluorescence. Antibodies generated against ZP1 in the same way were reactive with recombinant ZP proteins on Western blot only. The recombinant possum ZP proteins and specific antibodies produced in this study give an indication of the antigenic relationship of the possum ZP proteins and are vital tools for future studies of sperm\u2010ZP binding in marsupials and for the evaluation of ZP\u2010based contraceptive vaccines in possums and other marsupials. Mol. Reprod. Dev. 64: 136\u2013143, 2003. \u00a9 2003 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/mrd.10240","type":"journal-article","created":{"date-parts":[[2002,12,31]],"date-time":"2002-12-31T01:03:43Z","timestamp":1041296623000},"page":"136-143","source":"Crossref","is-referenced-by-count":15,"title":["Expression in <i>Escherichia coli<\/i> and immunological characterization of three zona pellucida proteins (ZP1, ZP2, and ZP3) from a marsupial, the brushtail possum (<i>Trichosurus vulpecula<\/i>)"],"prefix":"10.1002","volume":"64","author":[{"given":"Karen E.","family":"Mate","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Janine M.","family":"Buist","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Janine A.","family":"Duckworth","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2002,12,24]]},"reference":[{"key":"e_1_2_1_2_1","doi-asserted-by":"publisher","DOI":"10.1530\/rep.0.1230013"},{"key":"e_1_2_1_3_1","doi-asserted-by":"publisher","DOI":"10.1071\/R96058"},{"key":"e_1_2_1_4_1","unstructured":"DuckworthJ HarrisM MateK McCartneyC BuistJ ScobieS JonesD JonesS.1999.Development of brushtail possum immunocontraception: Targeting sperm and egg antigens. In Advances in the biological control of possums Vol. 56. The Royal Society of New Zealand Miscellaneous Series 72\u201376."},{"key":"e_1_2_1_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1071-5576(00)00107-6"},{"key":"e_1_2_1_6_1","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod57.3.532"},{"key":"e_1_2_1_7_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1098-2795(199902)52:2<174::AID-MRD8>3.0.CO;2-7"},{"key":"e_1_2_1_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1098-2795(199806)50:2<229::AID-MRD14>3.0.CO;2-1"},{"key":"e_1_2_1_9_1","doi-asserted-by":"publisher","DOI":"10.1530\/jrf.0.0950513"},{"key":"e_1_2_1_10_1","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod44.1.150"},{"key":"e_1_2_1_11_1","doi-asserted-by":"publisher","DOI":"10.1071\/RD01121"},{"key":"e_1_2_1_12_1","first-page":"159","article-title":"Influence of autoimmune disease on ZP3 contraceptive design","volume":"50","author":"Lou Y\u2010H","year":"1996","journal-title":"J Reprod Fertil"},{"key":"e_1_2_1_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0378-4320(98)00116-X"},{"key":"e_1_2_1_14_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1098-2795(199811)51:3<322::AID-MRD12>3.0.CO;2-4"},{"key":"e_1_2_1_15_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-642-58301-8_6"},{"key":"e_1_2_1_16_1","doi-asserted-by":"publisher","DOI":"10.1017\/S0967199400000976"},{"key":"e_1_2_1_17_1","doi-asserted-by":"publisher","DOI":"10.1017\/S0967199499000337"},{"key":"e_1_2_1_18_1","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod52.5.1167"},{"key":"e_1_2_1_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0303-7207(99)00236-1"},{"key":"e_1_2_1_20_1","doi-asserted-by":"publisher","DOI":"10.1530\/jrf.0.0760575"},{"key":"e_1_2_1_21_1","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod47.5.857"},{"key":"e_1_2_1_22_1","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod61.5.1356"},{"key":"e_1_2_1_23_1","doi-asserted-by":"publisher","DOI":"10.1017\/S0967199499000623"},{"key":"e_1_2_1_24_1","first-page":"189","volume-title":"The physiology of reproduction","author":"Yanagimachi R","year":"1994"}],"container-title":["Molecular Reproduction and Development"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/mrd.10240","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,11,17]],"date-time":"2023-11-17T11:36:24Z","timestamp":1700220984000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/mrd.10240"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2002,12,24]]},"references-count":23,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2003,2]]}},"alternative-id":["10.1002\/mrd.10240"],"URL":"https:\/\/doi.org\/10.1002\/mrd.10240","archive":["Portico"],"relation":{},"ISSN":["1040-452X","1098-2795"],"issn-type":[{"value":"1040-452X","type":"print"},{"value":"1098-2795","type":"electronic"}],"subject":[],"published":{"date-parts":[[2002,12,24]]}}}