<i>vp28</i> Gene Expression and Photosynthetic Characteristics of Transgenic <i>Synechococcus</i> sp. PCC 7942

doi:10.13523/j.cb.20180405

China Biotechnology

2018, Vol. 38

Issue (4): 30-37 DOI: 10.13523/j.cb.20180405

vp28 Gene Expression and Photosynthetic Characteristics of Transgenic Synechococcus sp. PCC 7942

Min-min ZHUANG¹,Xiao-hui JIA^1,²,Ding-ji SHI³(

),Jia-cheng ZHU¹,Si-yu FENG¹,Pei-min HE¹,Rui JIA¹(

)

1 School of Ocean Ecology and Environment Shanghai Ocean University, Shanghai 201306 China
2 School of Biosciences,Beijing Normal University,Beijing 100875, China
3 Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China

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Abstract

White spot syndrome virus (WSSV) is one of the most harmful viruses in the shrimp industry, and there has been no effective drug on mass scale up to now. In recent years, the immunological prevention of WSSV from shrimp has been progressed hopefully.Vp28 protein is the major structural component on the envelope of WSSV. Since 2004, its coding genes have been expressed in 8 species, and the control of WSSV has been proved remarkably in the laboratory. However, it has not been applied in shrimp industry yet. The shrimp bait, Synechococcus sp.PCC7942 was used as the acceptor to express vp28 gene, and this homology of medicine and food may be helpful for its application in the shrimp industry. The expression efficiency of vp28 in transgenic Synechococcus sp.PCC7942 has been detected by RT-qPCR method. And photosynthetic characteristics of transgenic Synechococcus sp.PCC7942 at different temperature, illumination, pH and salinity have been measured by the method of oxygen electrode. The expression efficiency of vp28 gene was 9.52% which was three times higher than that of Anabaena sp. PCC 7120. The most suitable harvest time was in late logarithm growth (the 15 ^th d). The optimum growth conditions of transgenic Synechococcus sp.PCC7942 were as follows: the temperature 40℃, the salinity 0~0.1mol/L NaCl, pH 7.5, light intensity 450μmol/(m ²·s). These data may be useful for scale preparation of oral drug made from transgenic Synechococcus sp.PCC7942.

Key words： Transgene Synechococcus with vp28 gene Photosynthetic activity RT-PCR Expression of vp28 gene

Received: 21 November 2017 Published: 08 May 2018

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	Min-min ZHUANG
	Xiao-hui JIA
	Ding-ji SHI
	Jia-cheng ZHU
	Si-yu FENG
	Pei-min HE
	Rui JIA

Cite this article:

Min-min ZHUANG,Xiao-hui JIA,Ding-ji SHI,Jia-cheng ZHU,Si-yu FENG,Pei-min HE,Rui JIA. vp28 Gene Expression and Photosynthetic Characteristics of Transgenic Synechococcus sp. PCC 7942. China Biotechnology, 2018, 38(4): 30-37.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20180405 OR https://manu60.magtech.com.cn/biotech/Y2018/V38/I4/30

Table 1 Cyanobacterial strains in this study

Table 2 vp28 primer sequence

Table 3 Real-time fluorescence quantitative PCR reaction conditions

Fig.1 Western blot of Vp28 from the wild type, blank, and transgenic type of Synechococcus sp.7942

Fig.2 Agarose gel electropherogram of vp28 PCR and melting curve of purified vp28 gene

Fig.3 Standard curve of recovered pure vp28 fragment

Table 4 Ct value and copy number of vp28 gene in transgenice Synechococcus sp.7942 at different growth stages

Table 5 Ct value and copy number of vp28 gene in transgenice Anabaena sp.7120 at different growth stages

Fig. 4 vp28 gene expression rates of transgenic Synechococcus sp.7942 during different growth stages

Fig.6 The responses of photosynthesis in the wild type and mutants of Synechococcus sp.7942 to environmental
(a) Photosynthesis-light curves (b) Photosynthesis - pH curves (c) Photosynthesis - temperature curves (d) Photosynthesis - salinity curves

Fig.7 Growth curves of wild type, empty vector type and transgenic type in Synechococcus sp.7942 type (detected at OD₇₅₀ of spectro photo meter)

Fig.8 Growth curves of wild type, empty vector type and transgenic type of Synechococcus sp.7942


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