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Green algal bestrophin bicarbonate transporters

A technology of bicarbonate and green algae, which is applied in the direction of algae/bryopeptide, microbe determination/inspection, peptide source, etc., and can solve the problem of few mechanism data available

Pending Publication Date: 2021-04-02
BOARD OF SUPERVISORS OF LOUISIANA STATE UNIV & AGRI & MECHANICAL COLLEGE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies now have identified many possible bicarbonate transporters, but only a few of these have promising experimental results and very little mechanistic data is available

Method used

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  • Green algal bestrophin bicarbonate transporters
  • Green algal bestrophin bicarbonate transporters
  • Green algal bestrophin bicarbonate transporters

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0132] Example 1: General Materials and Methods for Cell Culture and Growth

[0133] The following examples describe the cell culture and growth conditions used in all of the following examples. C. reinhardtii cells were maintained on triacetate phosphate (TAP) medium or yeast acetic acid (YA) medium on petri dishes. Before the experiment, cells were inoculated into minimal medium (i.e., without carbon source) and incubated under high CO 2 (>5% CO in air 2 ) grow to 2-3x10 6 cells mL -1 Density. These cells were then diluted into minimal medium and incubated at the indicated pH and CO 2 concentration growth.

[0134] C. reinhardtii culture conditions were the same as described in Ma et al., Plant Physiol 156:884-896, 2011. D66 strain (nit2 - , cw15, mt + ). CMJ030 (CC-4533; cwl5, mt - ) and bst3 (BST3 knockout LMJ.RY0402.089365). Triacetate phosphate (TAP) medium, acetic acid yeast (YA) medium and minimal medium (MIN; without acetate, ie without carbon source) were...

Embodiment 2

[0135] Embodiment 2: Phylogenetic tree of bestrophin family gene

[0136] The following examples describe the construction of phylogenetic trees depicting the evolutionary relationship of bestrophin family genes in various photosynthetic organisms. In mammals, bestrophin proteins are known to conduct chloride and bicarbonate. In plants, most bestrophin proteins have not been characterized.

[0137] Materials and methods

[0138] The three C. reinhardtii genes BESTROPHIN1 (BST1 ), BESTROPHIN2 (BST2 ) and BESTROPHIN3 (BST3 ) were chosen for use in this tree for several reasons. For one reason, the BST1, BST2, and BST3 genes share a common region (see Figures 1A-1B ). For another reason, the amino acid sequences of the BST1 (SEQ ID NO:1), BST2 (SEQ ID NO:2) and BST3 (SEQ ID NO:3) proteins are >80% identical to each other (see Figure 1C ). C. reinhardtii contains seven other predicted bestrophins, but none of these other bestrophins have greater than 45% identity to these ...

Embodiment 3

[0143] Example 3: At low CO 2 Lower Bestrophins are upregulated and their expression is controlled by CIA5

[0144] CIA5 is a transcription factor that controls many CCM genes. Specifically, CIA5 controls all known CCM transporters. The following examples describe the high CO 2 or low CO 2 Analysis of bestrophin expression in wild-type (WT) D66 C. reinhardtii strains and cia5 mutant C. reinhardtii strains grown under conditions.

[0145] Materials and methods

[0146] Cell culture and growth conditions for C. reinhardtii cells were as described in Example 1. For experiments, the WT D66 C. reinhardtii strain and the cia5 mutant C. reinhardtii strain were 2 (5% [v / v] CO in air 2 ) or low CO 2 Conditions (2 , that is, ambient CO 2 conditions) were cultivated.

[0147] RNA samples from two Chlamydomonas reinhardtii strains in high CO 2 and low CO 2 Adapted cultures were obtained. RNA extraction was performed using Trizol reagent (Invitrogen) according to the manufactu...

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Abstract

Aspects of the present disclosure relate to genetically modified plants and / or algae with increased carbon use efficiency as a result of an increased ability for bicarbonate to cross membranes withinplant cells. Other aspects of the present disclosure relate to methods of making such plants and / or algae as well as cultivating these genetically modified plants to increase carbon use efficiency and / or growing these genetically modified algae to increase carbon use efficiency.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of U.S. Provisional Application No. 62 / 769,214, filed November 19, 2018, and U.S. Provisional Application No. 62 / 697,840, filed July 13, 2018, which are hereby incorporated by reference in their entirety . [0003] Submission of a sequence listing as an ASCII text file [0004] The following submissions on the ASCII text file are hereby incorporated by reference in their entirety: Computer Readable Format of the Sequence Listing (CRF) (File Name: 794542000140SEQLIST.TXT, Date of Record: June 28, 2019, Size: 461KB). technical field [0005] The present disclosure relates to genetically modified plants. In particular, the present disclosure relates to genetically modified plants containing the bestrophin bicarbonate transporter of green algae, preferably providing increased carbon utilization efficiency. Background technique [0006] Chlorella and other photosynthetic aquatic organ...

Claims

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Application Information

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IPC IPC(8): A01H5/00A01H6/54A01H6/46A01H6/38C12N15/82C07K14/405
CPCC07K14/415C07K14/405C12N15/8269C12N15/8261C12N15/8218C12Q1/6895C12Q2600/13
Inventor A·慕克吉詹姆斯·V·莫罗尼C·S·刘L·C·M·麦金德
Owner BOARD OF SUPERVISORS OF LOUISIANA STATE UNIV & AGRI & MECHANICAL COLLEGE
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