A kind of corn sucrose synthase mutant gene, encoded protein and application thereof
A technology of sucrose synthase and protein, applied in the field of biogenetics, can solve the problem that the expression function of SuSy gene promoter is not very clear
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Embodiment 1
[0026] Example 1 Obtaining of sh1-m mutant
[0027] Obtaining the mutant: The sh1-m mutant was found in the material of the improved Zheng 58 inbred line (Gai-Z58). This protocol was obtained from an ear of the improved Zheng 58 inbred line in the experimental field of Henan Agricultural University in 2012. It was found that the endosperm was sunken in the grain, and the genetic stability of the trait was found through self-crossing experiments, and the endosperm of the mature grain shrunk spontaneously. Therefore, it was tentatively named sh1-m and further research was carried out.
[0028] In this program, the genetic control of the endosperm shrinkage phenotype was verified by crossing the mutant with the wild-type B73 and Mo17 varieties. The experimental results are shown in Table 1 below.
[0029] The ORF nucleotide sequence of the wild-type B73 is shown in SEQ ID No: 5, and the amino acid sequence of the protein encoded by the wild-type B73 gene is shown in SEQ ID No: 6....
Embodiment 2
[0034] The image clone of embodiment 2 sh1-m
[0035] (1) Preliminary positioning
[0036]Using 512 pairs of SSR primers synthesized in the laboratory, 5 polymorphic molecular markers P1-P5 linked to the target gene were found by means of BSA segregation analysis method, located in the Bin9.01 / 9.02 region (see Table 2 below). Use these 5 polymorphisms to mark 255 recessive BCs paired with B73 and sh1-m 1 Individuals were genotyped, and 39, 10, 12, 20 and 27 exchanged individuals were found, respectively. Marker P1 is located at the end close to the telomere, and the other 4 markers are located at the end close to the centromere, positioning the target gene within a fragment with a physical distance of about 8.3Mb between P1 and P2 (see figure 1 Middle A).
[0037] In order to further locate the sh1-m gene, nine polymorphic markers were newly identified, among which P6-P11 and P13-P14 were SSR markers, and P12 was an InDel marker (see Table 2 below). The 49 exchanged indivi...
Embodiment 3
[0048] Example 3 Allelism Test
[0049] In order to verify whether sh1-m is the allele of Sh1, two-way hybridization of sh1-m with sh1-912A was performed to obtain F 1 , by observing the mutants sh1-m and sh1-912A and their two F 1 Seeds spontaneously exhibit shrunken endosperm at maturity (see figure 2 ). This result confirmed that sh1-m is an allele of the Sh1 locus.
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