1. Expression Studies of Synthetic Sucrose Isomerase Gene 1 (SySIG1) in Saccharum officinarum L.
- Author
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Awan, Mudassar Fareed, Ali, Sajed, Ullah, Irfan, Ullah, Amin, Tariq, Muhammad, Iqbal, Muhammad Shahzad, Khalid, Aisha, Fatima, Sana, Shafiq, Yousuf, Nasir, Idrees Ahmad, Muzammal, Muhammad, and Farid, Arshad
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GENE expression ,SUCROSE ,TRANSGENE expression ,ISOMERS ,GENES ,GENE enhancers ,SUGARCANE ,ISOMERASES - Abstract
Extracted cane juice from Saccharum officinarum L. was employed as a major source of sugar acting as major input in various food industries. Current genotypes are yielding lower quantities of sugar posing a major challenge for sugarcane producers and plant biotechnologists. Discovery of novel sucrose isomerases (SI) derived from multiple microbial sources enlightened a great hope for molecular food scientists to introduce slowly digestible sucrose isomers not only to make up deficiencies of reduced sugar recovery but also produce healthful sugars. Modified Synthetic Sucrose Isomerase Gene 1 (SySIG1) cloned in plant expression vector pCAMBIA1301 under the combined influence of two promoters (CmYMV and Zm-Ubi) to produce high sugar yielding sugarcane genotypes. The expression of the transgene was terminated finally by nopaline synthase gene (Nos). The genetic construct pCEMBIA1301-1 was archestrated with transgene SySIGI having size 2180 bp integrated to plant expression vector. The vacoule targeted sequence (VTS) as signal peptide was also added in the vector so that the product could be directed and stored in the vacoular region. Transgene expression regulation was increased with enhancers to clone highly expressed gene construct. Transgene incorporation into genome was confirmed by PCR amplifications and dot blot analysis. Stem expression profile via qPCR indicated multiple fold magnification in transgene expression which was translated gorgeously into increased sugar recovery percentages in various tested transgenic lines comparative to control counterparts. Further quantifications by HPLC showed much high isomaltulose production in transgenic sugarcane lines as compared to control non-transgenic sugarcane lines. Different categories of sugarcane lines on the basis of sugar yields were developed and classified as super sugar lines (SSLs), elite sugar lines (ESLs), and good sugar lines (GSLs). Stem-based transgene expression and leaf-based expression quantifications were originated from real-time PCR. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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