Immune protection of Streptococcus iniae M-like protein mutant strain BY-1 ΔsimA

doi:10.3969/j.issn.1673-8640.2017.07.021

Abstract

Abstract:

Objective To investigate the immune protection of Streptococcus iniae M-like protein mutant strain BY-1 ΔsimA being used as a live attenuated vaccine to protect mice from Streptococcus iniae infection. Methods Allelic exchange mutagenesis was performed to construct Streptococcus iniae M-like protein mutant strain BY-1 ΔsimA. Shuttle vector pVE6007,which was sensitive to temperature,was used and could replicate in Streptococcus,to screen mutant strain BY-1 ΔsimA in a high efficiency. Kunming mice were immunized by the verified mutant strain BY-1 ΔsimA by intra-peritoneal injection,and infected with lethal dose Streptococcus iniae BY-1 after 21 d. The observation lasted for 15 d,and the accumulated death number was counted. Results Only 13.3% mice immunized with Streptococcus iniae mutant strain BY-1 ΔsimA were dead,resulting in 77.8% relative percentage survival（RPS）,which showed no statistical significance for wild type BY-1 whole-cell killed vaccine（P>0.05）,which had 55.5% of RPS. Conclusions Streptococcus iniae mutant strain BY-1 ΔsimA could effectively protect mice from Streptococcus iniae infection,which was an ideal live attenuated vaccine candidate.

Key words: BY-1ΔsimA, Relative percentage survival, Streptococcus iniae, Kunming mice

CLC Number:

R446.5

YANG Chengliang, LI Congrong. Immune protection of Streptococcus iniae M-like protein mutant strain BY-1 ΔsimA[J]. Laboratory Medicine, 2017, 32(7): 647-651.

Figures/Tables 4

References 21

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引物	序列（5'~3' ）
simA_F	CGCCATATGGATAGTGATAGATTAACCTTGGAAGNdeI
simA_R	CGCCTCGAGTTCTTCCTCTTTGCGTTTACGXhoI
simA_up_F	CGCGTCGACAGTCTGTTTCAAACTTGTCATGSalI
simA_up_R	TGTGTTTTATATTTTTCTCGTTCATGTTTAGGGTTCTCCTTATTTTC
simA_down_F	AATAGCTATAAATTATTTAATAAGTAAGCTTTCCCTTGCAACCTTTT
simA_down_R	CGCGGATCCAAGTACAAGGATGCTAGCCCTGBamHI
pVE6007_F	TAATACGACTCACTATAGGG
pVE6007_R	TCACTCATCATGTTCATATTTATCA
erm_F	GGAGTAACATAAATGAACGAGAAAAATATAAAACACAG
erm_R	CGTATCTTTCACTTATAAGAATATTACTTATTAAATAATTTATAGC
ITS_F	GAAGATAGGAAAGAGACGCAGTGTC
ITS_R	CCTTATTTCCAGTCTTTCGACCTTC

引物	序列（5'~3' ）
simA_F	CGCCATATGGATAGTGATAGATTAACCTTGGAAGNdeI
simA_R	CGCCTCGAGTTCTTCCTCTTTGCGTTTACGXhoI
simA_up_F	CGCGTCGACAGTCTGTTTCAAACTTGTCATGSalI
simA_up_R	TGTGTTTTATATTTTTCTCGTTCATGTTTAGGGTTCTCCTTATTTTC
simA_down_F	AATAGCTATAAATTATTTAATAAGTAAGCTTTCCCTTGCAACCTTTT
simA_down_R	CGCGGATCCAAGTACAAGGATGCTAGCCCTGBamHI
pVE6007_F	TAATACGACTCACTATAGGG
pVE6007_R	TCACTCATCATGTTCATATTTATCA
erm_F	GGAGTAACATAAATGAACGAGAAAAATATAAAACACAG
erm_R	CGTATCTTTCACTTATAAGAATATTACTTATTAAATAATTTATAGC
ITS_F	GAAGATAGGAAAGAGACGCAGTGTC
ITS_R	CCTTATTTCCAGTCTTTCGACCTTC