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As a way to circumvent this problem, we tested two different strategies for the development of synthetic promoters that are functionally equivalent but have a minimum sequence homology. Such promoters can be generated by (a) introducing known cis-elements in a novel or synthetic stretch of DNA or (b) \u201cdomain swapping,\u201d wherein domains of one promoter can be replaced with functionally equivalent domains from other heterologous promoters. We evaluated the two strategies for promoter modifications using domain A (consisting of minimal promoter and subdomain A1) of the 35S promoter as a model. A set of modified 35S promoters were developed whose strength was compared with the 35S promoter per se using \u03b2-glucuronidase as the reporter gene. Analysis of the expression of the reporter gene in transient assay system showed that domain swapping led to a significant fall in promoter activity. In contrast, promoters developed by placing cis-elements in a novel DNA context showed levels of expression comparable with that of the 35S. Two promoter constructs Mod2A1T and Mod3A1T were then designed by placing the core sequences of minimal promoter and subdomain A1 in divergent DNA sequences. Transgenics developed in tobacco (Nicotiana tabacum) with the two constructs and with 35S as control were used to assess the promoter activity in different tissues of primary transformants. Mod2A1T and Mod3A1T were found to be active in all of the tissues tested, at levels comparable with that of 35S. Further, the expression of the Mod2A1T promoter in the seedlings of the T1 generation was also similar to that of the 35S promoter. The present strategy opens up the possibility of creating a set of synthetic promoters with minimum sequence homology and with expression levels comparable with the wild-type prototype by modifying sequences present between cis-elements for transgene expression in plants.<\/jats:p>","DOI":"10.1104\/pp.103.020602","type":"journal-article","created":{"date-parts":[[2003,6,12]],"date-time":"2003-06-12T03:27:09Z","timestamp":1055388429000},"page":"988-998","source":"Crossref","is-referenced-by-count":87,"title":["Strategies for Development of Functionally Equivalent Promoters with Minimum Sequence Homology for Transgene Expression in Plants: cis-Elements in a Novel DNA Context versus Domain Swapping"],"prefix":"10.1093","volume":"132","author":[{"given":"Simran","family":"Bhullar","sequence":"first","affiliation":[{"name":"Department of Genetics, University of Delhi South Campus, Benito Juarez Road, New Delhi\u2013110021, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Suma","family":"Chakravarthy","sequence":"additional","affiliation":[{"name":"Department of Genetics, University of Delhi South Campus, Benito Juarez Road, New Delhi\u2013110021, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sonia","family":"Advani","sequence":"additional","affiliation":[{"name":"Department of Genetics, University of Delhi South Campus, Benito Juarez Road, New Delhi\u2013110021, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sudipta","family":"Datta","sequence":"additional","affiliation":[{"name":"Department of Genetics, University of Delhi South Campus, Benito Juarez Road, New Delhi\u2013110021, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deepak","family":"Pental","sequence":"additional","affiliation":[{"name":"Department of Genetics, University of Delhi South Campus, Benito Juarez Road, New Delhi\u2013110021, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pradeep Kumar","family":"Burma","sequence":"additional","affiliation":[{"name":"Department of Genetics, University of Delhi South Campus, Benito Juarez Road, New Delhi\u2013110021, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2003,6,1]]},"reference":[{"key":"2021042814480491800_REF1","doi-asserted-by":"crossref","unstructured":"An G, Ebert PR, Yi B-Y, Choi C-H (1986) Both TATA box and upstream regions are required for the nopaline synthase promoter activity in transformed tobacco cells. 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