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Table 1 Primers used for gene cloning and expression

From: A novel eurythermic and thermostale lipase LipM from Pseudomonas moraviensis M9 and its application in the partial hydrolysis of algal oil

Primers

Function

Sequence (5'-3')

T5

Degenerate primer

ACAATGTGCTGAACATNGGNTAYGARA

T3

Degenerate primer

GAAGGCGCCGCTGAANANNWANGTNTT

lipM9-5-1

Upstream genome walking

GACGCTTCCGCCAGGTTG

lipM9-5-2

Upstream genome walking

AGCCGTGAGCCGACCAGTTC

lipM9-5-3

Upstream genome walking

CCTGGTGTCCGTCGTGCTTG

lipM9-3-1

Downstream genome walking

GCTGTACGTGCGTGATGCCTAT

lipM9-3-2

Downstream genome walking

GGTGGGGGAGCAAGGAGGT

lipM9-3-3

Downstream genome walking

ACGGCAACACACTGGCAGC

lipM9-F

Complete genetic sequence

ATGGGAYTGTTCGATTACARAAAYGCCGA

lipM9-R

Complete genetic sequence

TYASGCRAASRYGAWRCYYGAAKCCGACAGG

lipM9-NF

Expression vector construction

CTCATATGGGATTGTTCGATTAC

lipM9-XR

Expression vector construction

CTCTCGAGCGCAAACATGATACTT

  1. a): Note: N represents A, C, G, or T; Y identifies C or T; R represents A or G; W identifies A or T. b): Nde I, Xho I restriction sites in primers are underlined, respectively