A broad absorption spectrum recombinant photosynthetic protein molecule and its construction method
A technology of absorption spectrum and photosynthetic protein, which is applied in the field of wide absorption spectrum recombination of photosynthetic protein molecules and its construction, can solve the problems of narrow absorption spectrum range and achieve broad absorption spectrum range, wide absorption range and wide sources Effect
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Embodiment 1
[0039] PCR amplification of target gene
[0040]Using pUC19-LhcⅡ as template LHCⅡ-F and LHCⅡ-PCA-R as primers, pCDFDuet-cpcA-cpcEF-hox1-pcyA plasmid as template LHCⅡ-PCA-F and PCA-R as primers to amplify LhcⅡ and cpcA genes respectively, The theoretical values of the target bands are 687bp and 492bp respectively. detected by electrophoresis as figure 1 As shown, referring to the Marker as D2000, it is found that the bands are clear under ultraviolet irradiation. The actual size of the LHC II gene is basically consistent with the theoretical value at the position slightly lower than the third band of Marker at 750bp, and the actual size of the PCA gene is in the fourth band of the Marker. The band 500bp is basically consistent with the theoretical value, figure 1 Among them, a is LhcⅡ fragment; b is Marker; c is cpcA fragment.
Embodiment 2
[0042] Overlap PCR
[0043] The LhcII and cpcA genes recovered from the gel were used as templates for overlapping PCR to connect the two fragments. The size of the LHCⅡ-PCA fusion fragment is 1200bp. After electrophoresis detection, the reference marker is D2000. It is found that the size of the target band is slightly above 1000bp, which is basically consistent with the theoretical value of 1200bp, which proves that the fusion fragment is connected successfully. The experimental results are as follows figure 2 as shown, figure 2 Among them, a is the LhcⅡ-cpcA fragment; b is Marker:
Embodiment 3
[0045] double digestion
[0046] Using Marker (1kb) as a reference, the plasmid vectors pCDFDuet-cpcA-cpcEF-hox1-pcyA and pCDFDuet-cpcA-cpcEF-hox1-pebS after BamHI and SacⅠ double digestion, can see clear large molecular weight target band fragments, And it is basically consistent with the theoretical value of 6kb, and there are excised small gene fragments at the lower molecular weight, the results are as follows image 3 as shown, image 3 Among them, a is the carrier containing PEB; b is the excised small gene fragment; c is the carrier containing PCB; d is the excised small gene fragment:
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