{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,7]],"date-time":"2026-02-07T22:26:38Z","timestamp":1770503198415,"version":"3.49.0"},"reference-count":27,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"4987","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[1990,12,14]]},"abstract":"<jats:p>\n            Development of methods for the manipulation of the genomes of parasitic protozoa will lead to enhanced understanding of parasite biology and host-parasite relationships. Efficient gene transfer and targeted integration by homologous recombination were achieved in the parasitic protozoan\n            <jats:italic>Trypanosoma brucei<\/jats:italic>\n            , the causative agent of sleeping sickness. An expression vector with the neomycin phosphotransferase gene (\n            <jats:italic>neo<\/jats:italic>\n            ), under the control of a procyclic acidic repetitive protein (PARP) gene promoter, was targeted into an intergenic region in \u03b2\u03b1-tubulin-gene tandem array. Sixteen copies of\n            <jats:italic>neo<\/jats:italic>\n            were found in a tandem array in one of the transfectants where the PARP promoter controlled \u03b1-amanitin-resistant transcription of\n            <jats:italic>neo<\/jats:italic>\n            , whereas transcription of tubulin genes remained \u03b1-amanitin-sensitive.\n          <\/jats:p>","DOI":"10.1126\/science.2177225","type":"journal-article","created":{"date-parts":[[2006,10,5]],"date-time":"2006-10-05T22:10:14Z","timestamp":1160086214000},"page":"1583-1587","source":"Crossref","is-referenced-by-count":99,"title":["Homologous Recombination and Stable Transfection in the Parasitic Protozoan\n            <i>Trypanosoma brucei<\/i>"],"prefix":"10.1126","volume":"250","author":[{"given":"Mary Gwo-Shu","family":"Lee","sequence":"first","affiliation":[]},{"given":"Lex H. T.","family":"Van der Ploeg","sequence":"additional","affiliation":[]}],"member":"221","reference":[{"key":"e_1_2_1_1_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.2499047"},{"key":"e_1_2_1_2_1","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1146\/annurev.bi.55.070186.003413","volume":"55","year":"1986","unstructured":"BORST, P, DISCONTINUOUS TRANSCRIPTION AND ANTIGENIC VARIATION IN TRYPANOSOMES, ANNUAL REVIEW OF BIOCHEMISTRY 55: 701 (1986).","journal-title":"ANNUAL REVIEW OF BIOCHEMISTRY"},{"key":"e_1_2_1_3_1","first-page":"3036","volume":"10","year":"1990","unstructured":"CLAYTON, C.E., TRANSCRIPTION OF THE PROCYCLIC ACIDIC REPETITIVE PROTEIN GENES OF TRYPANOSOMA-BRUCEI, MOLECULAR AND CELLULAR BIOLOGY 10: 3036 (1990).","journal-title":"MOLECULAR AND CELLULAR BIOLOGY"},{"key":"e_1_2_1_4_1","first-page":"4657","volume":"6","year":"1986","unstructured":"GLASS, D.J., CONSERVED SEQUENCES AND TRANSCRIPTION OF THE HSP70-GENE FAMILY IN TRYPANOSOMA-BRUCEI, MOLECULAR AND CELLULAR BIOLOGY 6: 4657 (1986).","journal-title":"MOLECULAR AND CELLULAR BIOLOGY"},{"key":"e_1_2_1_5_1","first-page":"6079","volume":"10","year":"1990","unstructured":"GOTTESDIENER, K, CHROMOSOME ORGANIZATION OF THE PROTOZOAN TRYPANOSOMA-BRUCEI, MOLECULAR AND CELLULAR BIOLOGY 10: 6079 (1990).","journal-title":"MOLECULAR AND CELLULAR BIOLOGY"},{"key":"e_1_2_1_6_1","first-page":"1084","volume":"10","year":"1990","unstructured":"KAPLER, G.M., STABLE TRANSFECTION OF THE HUMAN PARASITE LEISHMANIA-MAJOR DELINEATES A 30-KILOBASE REGION SUFFICIENT FOR EXTRACHROMOSOMAL REPLICATION AND EXPRESSION, MOLECULAR AND CELLULAR BIOLOGY 10: 1084 (1990).","journal-title":"MOLECULAR AND CELLULAR BIOLOGY"},{"key":"e_1_2_1_7_1","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1016\/0092-8674(87)90153-X","volume":"51","year":"1987","unstructured":"KOOTER, J.M., THE ANATOMY AND TRANSCRIPTION OF A TELOMERIC EXPRESSION SITE FOR VARIANT-SPECIFIC SURFACE-ANTIGENS IN TRYPANOSOMA-BRUCEI, CELL 51: 261 (1987).","journal-title":"CELL"},{"key":"e_1_2_1_8_1","doi-asserted-by":"crossref","first-page":"9457","DOI":"10.1093\/nar\/12.24.9457","volume":"12","year":"1984","unstructured":"KOOTER, J.M., ALPHA-AMANITINE-INSENSITIVE TRANSCRIPTION OF VARIANT SURFACE GLYCOPROTEIN GENES PROVIDES FURTHER EVIDENCE FOR DISCONTINUOUS TRANSCRIPTION IN TRYPANOSOMES, NUCLEIC ACIDS RESEARCH 12: 9457 (1984).","journal-title":"NUCLEIC ACIDS RESEARCH"},{"key":"e_1_2_1_9_1","doi-asserted-by":"crossref","first-page":"572","DOI":"10.1038\/343572a0","volume":"343","year":"1990","unstructured":"LABAN, A, STABLE EXPRESSION OF THE BACTERIAL NEOR GENE IN LEISHMANIA-ENRIETTII, NATURE 343: 572 (1990).","journal-title":"NATURE"},{"key":"e_1_2_1_10_1","first-page":"9119","volume":"86","year":"1989","unstructured":"LABAN, A, TRANSFECTION OF LEISHMANIA-ENRIETTII AND EXPRESSION OF CHLORAMPHENICOL ACETYLTRANSFERASE GENE, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 86: 9119 (1989).","journal-title":"PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA"},{"key":"e_1_2_1_11_1","doi-asserted-by":"crossref","first-page":"4253","DOI":"10.1093\/nar\/13.12.4253","volume":"13","year":"1985","unstructured":"LAIRD, P.W., MATURE MESSENGER-RNAS OF TRYPANOSOMA-BRUCEI POSSESS A 5' CAP ACQUIRED BY DISCONTINUOUS RNA-SYNTHESIS, NUCLEIC ACIDS RESEARCH 13: 4253 (1985).","journal-title":"NUCLEIC ACIDS RESEARCH"},{"key":"e_1_2_1_12_1","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/0166-6851(90)90184-N","volume":"41","year":"1990","unstructured":"LEE, MGS, EVIDENCE FOR SEGMENTAL GENE CONVERSION BETWEEN A COGNATE HSP70 GENE AND THE TEMPERATURE-SENSITIVELY TRANSCRIBED HSP70 GENES OF TRYPANOSOMA-BRUCEI, MOLECULAR AND BIOCHEMICAL PARASITOLOGY 41: 213 (1990).","journal-title":"MOLECULAR AND BIOCHEMICAL PARASITOLOGY"},{"key":"e_1_2_1_13_1","doi-asserted-by":"crossref","first-page":"9567","DOI":"10.1093\/nar\/16.20.9567","volume":"16","year":"1988","unstructured":"LEE, MGS, STRUCTURE AND EXPRESSION OF THE HSP-70 GENE FAMILY OF LEISHMANIA-MAJOR, NUCLEIC ACIDS RESEARCH 16: 9567 (1988).","journal-title":"NUCLEIC ACIDS RESEARCH"},{"key":"e_1_2_1_14_1","unstructured":"LEE MGS unpublished data."},{"key":"e_1_2_1_15_1","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1128\/mr.49.1.33-58.1985","volume":"49","year":"1985","unstructured":"ORRWEAVER, T.L., FUNGAL RECOMBINATION, MICROBIOLOGICAL REVIEWS 49: 33 (1985).","journal-title":"MICROBIOLOGICAL REVIEWS"},{"key":"e_1_2_1_16_1","first-page":"6354","volume":"78","year":"1981","unstructured":"ORRWEAVER, T.L., YEAST TRANSFORMATION - A MODEL SYSTEM FOR THE STUDY OF RECOMBINATION, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES 78: 6354 (1981).","journal-title":"PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES"},{"key":"e_1_2_1_17_1","doi-asserted-by":"crossref","first-page":"4259","DOI":"10.1002\/j.1460-2075.1989.tb08611.x","volume":"8","year":"1989","unstructured":"RUDENKO, G, EMBO JOURNAL 8: 4259 (1989).","journal-title":"EMBO JOURNAL"},{"key":"e_1_2_1_18_1","first-page":"3492","volume":"10","year":"1990","unstructured":"RUDENKO, G, PROCYCLIC ACIDIC REPETITIVE PROTEIN (PARP) GENES LOCATED IN AN UNUSUALLY SMALL ALPHA-AMANITIN-RESISTANT TRANSCRIPTION UNIT - PARP PROMOTER ACTIVITY ASSAYED BY TRANSIENT DNA TRANSFECTION OF TRYPANOSOMA-BRUCEI, MOLECULAR AND CELLULAR BIOLOGY 10: 3492 (1990).","journal-title":"MOLECULAR AND CELLULAR BIOLOGY"},{"key":"e_1_2_1_19_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.2448875"},{"key":"e_1_2_1_20_1","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/0022-2836(80)90196-5","volume":"143","year":"1980","unstructured":"SANGER, F, CLONING IN SINGLE-STRANDED BACTERIOPHAGE AS AN AID TO RAPID DNA SEQUENCING, JOURNAL OF MOLECULAR BIOLOGY 143: 161 (1980).","journal-title":"JOURNAL OF MOLECULAR BIOLOGY"},{"key":"e_1_2_1_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2836(75)80083-0"},{"key":"e_1_2_1_22_1","first-page":"327","volume":"1","year":"1982","unstructured":"SOUTHERN, P.J., JOURNAL OF MOLECULAR AND APPLIED GENETICS 1: 327 (1982).","journal-title":"JOURNAL OF MOLECULAR AND APPLIED GENETICS"},{"key":"e_1_2_1_23_1","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/0092-8674(83)90331-8","volume":"33","year":"1983","unstructured":"SZOSTAK, J.W., THE DOUBLE-STRAND-BREAK REPAIR MODEL FOR RECOMBINATION, CELL 33: 25 (1983).","journal-title":"CELL"},{"key":"e_1_2_1_24_1","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/0092-8674(83)90494-4","volume":"32","year":"1983","unstructured":"THOMASHOW, L.S., TUBULIN GENES ARE TANDEMLY LINKED AND CLUSTERED IN THE GENOME OF TRYPANOSOMA BRUCEI, CELL 32: 35 (1983).","journal-title":"CELL"},{"key":"e_1_2_1_25_1","first-page":"51","year":"1990","unstructured":"Van der Ploeg, L. H. T., Frontiers in Molecular Biology; Genome Rearrangements and Amplification: 51 (1990).","journal-title":"Frontiers in Molecular Biology; Genome Rearrangements and Amplification"},{"key":"e_1_2_1_26_1","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1093\/nar\/10.2.593","volume":"10","year":"1982","unstructured":"VANDERPLOEG, LHT, CHARACTERIZATION OF THE DNA DUPLICATION-TRANSPOSITION THAT CONTROLS THE EXPRESSION OF 2 GENES FOR VARIANT SURFACE GLYCOPROTEINS IN TRYPANOSOMA-BRUCEI, NUCLEIC ACIDS RESEARCH 10: 593 (1982).","journal-title":"NUCLEIC ACIDS RESEARCH"},{"key":"e_1_2_1_27_1","doi-asserted-by":"crossref","first-page":"3217","DOI":"10.1093\/nar\/17.8.3217","volume":"17","year":"1989","unstructured":"VANDERPLOEG, LHT, IMPROVED SEPARATION OF CHROMOSOME-SIZED DNA FROM TRYPANOSOMA-BRUCEI, STOCK-427-60, NUCLEIC ACIDS RESEARCH 17: 3217 (1989).","journal-title":"NUCLEIC ACIDS RESEARCH"}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.2177225","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,12]],"date-time":"2024-01-12T09:42:48Z","timestamp":1705052568000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.2177225"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1990,12,14]]},"references-count":27,"journal-issue":{"issue":"4987","published-print":{"date-parts":[[1990,12,14]]}},"alternative-id":["10.1126\/science.2177225"],"URL":"https:\/\/doi.org\/10.1126\/science.2177225","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[1990,12,14]]}}}