Scholarlink

<p>Constitutive gene expression can lead to pleiotropic effects. Therefore, spatial or temporal restriction of expression via specific promoters provides a more targeted approach. This study aimed to clone the <i>SalT</i> promoter and analyze its activity in transgenic barley using <i>GFP</i> and <i>GUS</i> reporter genes. The T-DNA constructs carrying the <i>SalT</i> promoter were introduced into barley cv. Golden Promise, and transgenic plants were confirmed through PCR, hygromycin selection, and Southern hybridization. Both constructs, <i>Sal</i>T-GFP and <i>Sal</i>T-GUS, were transformed in barley cv. Golden Promise. Here, we characterized the expression pattern of the <i>Sal</i>T promoter in barley and utilized it to drive the expression of reporter genes <i>GFP</i> and <i>GUS</i>. The <i>SalT</i> promoter was isolated from rice genomic DNA, cloned into the pNos-AB-M vector, and confirmed through PCR and restriction analysis. Subsequently, <i>GFP</i> and <i>GUS</i> genes were cloned under the <i>SalT</i> promoter in the same vector. The constructs were then subcloned into the p6U vector for plant expression. <i>Agrobacterium</i>-mediated genetic transformation of barley cultivar “Golden Promise" was conducted, resulting in successful integration of the transgenes. Callus induction, regeneration, and root formation efficiency were assessed, demonstrating the potential of the <i>SalT</i> promoter to drive gene expression during various stages of plant development. Molecular analyses, including PCR and Southern hybridization, confirmed the presence and integration of transgenes in the barley genome. Furthermore, GFP fluorescence and GUS staining analyses revealed strong expression of the respective genes under control of the <i>SalT</i> promoter in different plant tissues. This study provides insights into the application of the <i>SalT</i> promoter for genetic manipulation and functional characterization in barley, offering opportunities for crop improvement and biotechnological applications.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Cloning, Transformation, and Reporter Gene Analysis of the SalT Promoter in Barley (Hordeum vulgare)

  • 期刊论文

  • 发表日期：

    2026年5月1日

查看全文
  • Zahid Abbas Malik,
  • Jochen Kumlehn,
  • Sabir Hussain Shah,
  • Zeshan Hassan,
  • Götz Hensel,
  • Nasir Ahmad Saeed

摘要

Constitutive gene expression can lead to pleiotropic effects. Therefore, spatial or temporal restriction of expression via specific promoters provides a more targeted approach. This study aimed to clone the SalT promoter and analyze its activity in transgenic barley using GFP and GUS reporter genes. The T-DNA constructs carrying the SalT promoter were introduced into barley cv. Golden Promise, and transgenic plants were confirmed through PCR, hygromycin selection, and Southern hybridization. Both constructs, SalT-GFP and SalT-GUS, were transformed in barley cv. Golden Promise. Here, we characterized the expression pattern of the SalT promoter in barley and utilized it to drive the expression of reporter genes GFP and GUS. The SalT promoter was isolated from rice genomic DNA, cloned into the pNos-AB-M vector, and confirmed through PCR and restriction analysis. Subsequently, GFP and GUS genes were cloned under the SalT promoter in the same vector. The constructs were then subcloned into the p6U vector for plant expression. Agrobacterium-mediated genetic transformation of barley cultivar “Golden Promise" was conducted, resulting in successful integration of the transgenes. Callus induction, regeneration, and root formation efficiency were assessed, demonstrating the potential of the SalT promoter to drive gene expression during various stages of plant development. Molecular analyses, including PCR and Southern hybridization, confirmed the presence and integration of transgenes in the barley genome. Furthermore, GFP fluorescence and GUS staining analyses revealed strong expression of the respective genes under control of the SalT promoter in different plant tissues. This study provides insights into the application of the SalT promoter for genetic manipulation and functional characterization in barley, offering opportunities for crop improvement and biotechnological applications.