TAILIEUCHUNG - analysis of genes and genomes phần 6

miền của các Herpes simplex VP16 protein virus hoặc kích hoạt trình tự nhân tạo B42), xem xét bởi Brent và Finley (1997). Các nguyên tắc mô tả ở trên cho các hệ thống dựa trên Gal4p giữ đúng đối với mỗi người trong các phiên bản sửa đổi. | 228 GENE IDENTIFICATION 6 domain of the Herpes simplex virus protein VP16 or the artificial activating sequence B42 reviewed by Brent and Finley 1997 . The principles described above for the Gal4p based systems hold true for each of these modified versions. Other Interaction Screens - Variations on a Theme The two-hybrid screen has been modified extensively to suit different purposes. The basic objective behind each type of screen that is listed below is to assemble One-hybrid screen to identify DNA-binding domains b uasg Three-hybrid screen to identify RNA-binding proteins c Bifunctional ligand LexOp Three-hybrid screen to identify ligand-binding proteins Figure . Other types of interaction screening. a One-hybrid screening. b c Three-hybrid screening. See the text for details OTHER INTERACTION SCREENS - VARIATIONS ON A THEME 229 a functional transcriptional activator protein within the promoter of a reporter gene whose expression can then be detected Figure . One Hybrid In a simplification of the two-hybrid system a cDNA-AD library is introduced into a yeast strain that harbours a reporter gene into which DNA binding sites have been introduced. Proteins encoded by the cDNA that are able to bind to the DNA binding sites will activate the transcription of the reporter gene Li and Herskowitz 1993 . This approach is useful in the identification of proteins that regulate the promoters of known genes Wang and Reed 1993 . To perform a screen like this the DNA binding element is usually multimerized to produce a strong activation element through the binding of multiple activator proteins so that expression of the reporter can be observed easily. Three Hybrid Any mechanism by which the DBD and AD functions of the transcriptional activator may be brought together can be used to activate the reporter gene. Several screens have been described in which the DBD and AD fusion proteins do not interact directly with each other but their interaction is

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