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EXPRESSYALI
(Kit # 1000000245 )

Depositing Lab:   Irina Borodina

The EXPRESSYALI Set is a complete kit for the introduction of genes into Yarrowia chromosomal loci. The kit includes six part plasmids, 12 marker-free integrative vectors, and 11 gRNA plasmids.

This kit will be sent as bacterial glycerol stocks in 96-well plate format.

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$400 USD + shipping
Available to academics and nonprofits only.

Original Publication

Combinatorial iterative method for metabolic engineering of Yarrowia lipolytica: application for betanin biosynthesis. Jiang W, Wang S, Avila P, Jorgensen TS, Yang Z, Borodina I. Metab Eng. 2024 Sep 9:S1096-7176(24)00110-1. doi: 10.1016/j.ymben.2024.09.003. PubMed (Link opens in a new window) Article (Link opens in a new window).

Description

The EXPRESSYALI collection facilitates the hierarchical assembly of single and multi-gene constructs, up to three transcription units, for Yarrowia lipolytica expression. Developed by Irina Borodina’s lab, this toolkit includes three well-characterized promoters, three terminators, 12 marker-free integrative vectors, and 11 gRNA plasmids. It employs the Golden Gate cloning method to enable the generation of a combinatorial library from three distinct transcription units. Additionally, the toolkit utilizes two interchangeable selection markers—NatMX for nourseothricin and HphMX for hygromycin—in successive transformation rounds, eliminating the need for marker excision.

Bioparts can be obtained by DNA synthesis or by PCR amplification if the template DNA is already available. The Bioparts are cloned into the PCR-Blunt II TOPO vector using TOPO® cloning, resulting in the generation of part plasmids (Level 0). The part plasmids, including promoters, coding sequences, and terminators, are assembled into YALICP vectors to form single-cassette plasmids (Level 1) through the BsaI Golden Gate method. Single-cassette plasmids are further combined to construct multigene-cassette plasmids (Level 2) utilizing the BsmbI Golden Gate assembly method. A combinatorial library can be generated through random combinations of different Level 1 YALICP (a), (b), and (c) plasmids. After the combinatorial library is introduced into Y. lipolytica, the optimal clones can be identified using high-throughput screening technologies. By leveraging various integration sites, the library can be iteratively applied.
  • Figure 1. EXPRESSYALI system workflow consisted of iterative rounds of transformation of combinatorial libraries into yeast Y. lipolytica with selection using high-throughput technology. HR: homologous arm. Ori: the origin of replication. SP: spectinomycin. AMP: ampicillin.

How to Cite this Kit

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.

For your Materials and Methods section:

“The EXPRESSYALI was a gift from Irina Borodina (Addgene kit #1000000245).”

For your Reference section:

Combinatorial iterative method for metabolic engineering of Yarrowia lipolytica: application for betanin biosynthesis. Jiang W, Wang S, Avila P, Jorgensen TS, Yang Z, Borodina I. Metab Eng. 2024 Sep 9:S1096-7176(24)00110-1. doi: 10.1016/j.ymben.2024.09.003. PubMed (Link opens in a new window) Article (Link opens in a new window).

EXPRESSYALI - #1000000245

Resistance Color Key

Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.

Inventory

Searchable and sortable table of all plasmids in kit. The Well column lists the plasmid well location in its plate. The Plasmid column links to a plasmid's individual web page.

Kit Plate Map

96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.

Resistance Color Key

Ampicillin
Spectinomycin
Kanamycin
Chloramphenicol

Inventory

Well Plasmid Resistance
A / 1 pCfB12714
Ampicillin
A / 2 pCfB12715
Ampicillin
A / 3 pCfB12716
Ampicillin
A / 4 pCfB12717
Ampicillin
A / 5 pCfB12718
Ampicillin
A / 6 pCfB13017
Ampicillin
A / 7 pCfB10769
Ampicillin
A / 8 pCfB11787
Ampicillin
A / 9 pCfB6637
Ampicillin
A / 10 pCfB8860
Ampicillin
A / 11 pCfB6638
Ampicillin
A / 12 pCfB11785
Ampicillin
B / 1 pCfB6630
Ampicillin
B / 2 pCfB8856
Ampicillin
B / 3 pCfB12719
Ampicillin
B / 4 pCfB8866
Ampicillin
B / 5 pCfB12720
Ampicillin
B / 6 pCfB12711
Spectinomycin
B / 7 pCfB12712
Spectinomycin
B / 8 pCfB12713
Spectinomycin
B / 9 pCfB13014
Spectinomycin
B / 10 pCfB13015
Spectinomycin
B / 11 pCfB13016
Spectinomycin
B / 12 pCfB12706
Kanamycin
C / 1 pCfB12708
Kanamycin
C / 2 pCfB12710
Kanamycin
C / 3 PCfB12709
Kanamycin
C / 4 pCfB13089
Chloramphenicol
C / 5 pCfB13090
Chloramphenicol
Data calculated @ 2024-11-21

Kit Plate Map - #1000000245

Plate #1

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