title: MULTIPLEX PCR ASSAY FOR DETECTION OF HUMAN SOMATOTROPIN AND INTERFERON ALPHA2b GENES IN PLANT MATERIAL.
abstract: Using transgenic plants as factories for production of physiologically active human proteins arouses special concern because occasional escape of such transgenes into environment may cause health problems. Creation of plant varieties producing pharmaceutically valuable proteins should be accompanied by development of detection methods suitable for controlling the transgene behavior. Here we describe a multiplex PCR protocol for revealing of two human genes (encoding growth hormone and interferon alpha2b) that have been successfully introduced into plant genomes. The primer pair designed for detection of human growth hormone coding sequence amplifies fragments of different size from the full-length gene in the human genome and the intronless coding sequence usually used for plant transformation. Application of this primer pair may be recommended for ruling out false positive results due to sample contamination with human DNA. Such a control may be useful also in PCR analysis during establishing of transgenic plants carrying genes of human origin.
authors.forename: I M
authors.initials: IM
authors.lastname: Gerasymenko
authors.forename: M G
authors.initials: MG
authors.lastname: Mazur
authors.forename: Y V
authors.initials: YV
authors.lastname: Sheludko
authors.forename: N V
authors.initials: NV
authors.lastname: Kuchuk
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D048013:Acetivibrio thermocellus / Q000737:chemistry; D017931:DNA Primers / Q000138:chemical synthesis / Q000737:chemistry*; D015870:Gene Expression; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism; D013006:Growth Hormone / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics*; D014170:Transformation, Genetic; D019076:Transgenes
chemical_list: D017931:DNA Primers; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D013006:Growth Hormone; D006026:Glycoside Hydrolases; C028020:licheninase