Difference between revisions of "Part:BBa K4887001"
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| − | <b> (1) GBSSI-sgRNA design </ | + | <h1><b>Results</b></h1> |
| + | <h2>(1) GBSSI-sgRNA design</h2> | ||
The selected sgRNA is located 70 bp downstream of the start codon, with a total sequence length of 27 bp (Fig. 1). sgRNA oligos were obtained by annealing with the designed primers following: | The selected sgRNA is located 70 bp downstream of the start codon, with a total sequence length of 27 bp (Fig. 1). sgRNA oligos were obtained by annealing with the designed primers following: | ||
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| − | < | + | <h2>(2) The expression level of IbGBSSI in storage roots</h2> |
The relative expression level of the gene IbGBSSI was determined with the method of Quantitative Real-time PCR with the root-tuber cDNA as templates. The result showed that the relative expression level of IbGBSSI in root tubers of the transgenic lines (0.1063 and 0.2407) was much lower than that of the wild type (1.0000) (Fig. 2). It revealed that the knock-out of IbGBSSI in the pathway of starch synthesis was successful. | The relative expression level of the gene IbGBSSI was determined with the method of Quantitative Real-time PCR with the root-tuber cDNA as templates. The result showed that the relative expression level of IbGBSSI in root tubers of the transgenic lines (0.1063 and 0.2407) was much lower than that of the wild type (1.0000) (Fig. 2). It revealed that the knock-out of IbGBSSI in the pathway of starch synthesis was successful. | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K4887001 SequenceAndFeatures</partinfo> | <partinfo>BBa_K4887001 SequenceAndFeatures</partinfo> | ||
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<!-- Uncomment this to enable Functional Parameter display | <!-- Uncomment this to enable Functional Parameter display | ||
Revision as of 10:03, 6 October 2023
IbGBSSI
The encoding gene of granule-bound starch synthase I (GBSSI) in sweet potato (Ipomoea batatas). It is one of the starch biosynthetic pathway genes on the starch biosynthetic pathway.
Results
(1) GBSSI-sgRNA design
The selected sgRNA is located 70 bp downstream of the start codon, with a total sequence length of 27 bp (Fig. 1). sgRNA oligos were obtained by annealing with the designed primers following: Oligo 1 (GBSSI-sgRNA1): 5’-GTGGGGTTGGGTCAATTAGCCCTGAGGAGC-3' Oligo 2 (GBSSI-sgRNA2): 5’-AACTGGTGGACTTGGAGATGTTCTTGGAGG-3’
Fig. 1 The target region of gene IbGBSSI and the location of the gRNA. Exons are shown as square frames and surrounding introns appear as lines. sgRNA and PAM are highlighted in yellow and green, respectively.
(2) The expression level of IbGBSSI in storage roots
The relative expression level of the gene IbGBSSI was determined with the method of Quantitative Real-time PCR with the root-tuber cDNA as templates. The result showed that the relative expression level of IbGBSSI in root tubers of the transgenic lines (0.1063 and 0.2407) was much lower than that of the wild type (1.0000) (Fig. 2). It revealed that the knock-out of IbGBSSI in the pathway of starch synthesis was successful.
Fig. 2 Q-PCR result of the relative expression level of IbGBSSI in root tubers
Sequence and Features