Cloning and expression of the constant region of rainbow trout (Onchorhynchus mykiss) µ immunoglobulin chain in Escherichia coli

نویسندگان

چکیده مقاله:

The importance of rainbow trout (Onchorhynchus mykiss) in Iran aquaculture industry on one hand, and increasing the mortality of this fish due to outbreaks of infectious diseases, on the other hand, indicate the requirement for more profound understanding the rainbow trout immune system and access to laboratory tools for definitive diagnosis of its diseases. One of the most important defense mechanisms of vertebrates including fish is the production of immunoglobulin against microbial pathogens. In rainbow trout, dominant immunoglobulin in serum is immunoglobulin M (IgM). The purpose of this study was the cloning and expression of the constant region of rainbow trout IgM heavy chain (µ chain) gene in Escherichia coli. Therefore, RNA of the targeted gene was extracted from spleen and head kidney of rainbow trout and the constant region of µ chain was amplified by RT-PCR. The amplified fragment was ligated to pMALc2x vector and transferred to DH5α strain of E. coli. Recombinant vector transformed and expressed into E. coli Rosetta strain. SDS-PAGE analysis indicated the production of a recombinant protein with an expected molecular weight of 75 KDa. Thereafter, the recombinant protein was purified by amylose resin and its antigenicity was accessed by immunoblotting. Positive reaction of the expressed protein with anti-trout serum indicated that the expressed constant region of trout µ chain possess antigenic epitopes and could be applied in future immunological studies.

برای دانلود باید عضویت طلایی داشته باشید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Genetic Characterization of Streptococcus iniae in Diseased Farmed Rainbow Trout (Onchorhynchus mykiss) in Iran

Genetic characterization of strains of Streptococcus iniae recovered from morbidity and mortality of farmed rainbow trout in different provinces of Iran were studied. The Gram-positive cocci isolates were obtained from the kidney tissues of diseased rainbow trout on blood agar at 25°C for 72 h. The grown bacteria were then characterized using biochemical and molecular works. The identified 26 i...

متن کامل

Development of 16S rRNA targeted PCR methods for the detection of Escherichia coli in Rainbow trout (Oncorhynchus mykiss)

  Objectives: The presence of E.coli in fish intended for human consumption may constitute a potential danger, not only in causing disease, but also because of the possible transfer of antibiotic resistance from aquatic bacteria to those infecting humans. The objective of this study was to develop an improved PCR method based on species – specific 16 S rRNA gene primers (FES,...

متن کامل

The use of encapsulated oil in dietary on growth factors, proximate compositions and frozen storage of rainbow trout (Onchorhynchus mykiss)

This study was performed in order to survey of effect of encapsulated oil in rain bow trout dietary on some growth factors and proximate compositions. Fish in average weight, 20 ± 1.5 g, was fed daily at 2.5 percent of biomass for 2 months. Rainbow trout was weighed every 2 weeks. At the end of experiment growth factors and proximate composition were studied. The result showed that the diets wh...

متن کامل

Accumulation of 2,3,7,8-tetrachlorodibenzo-p-dioxin by rainbow trout (Onchorhynchus mykiss) at environmentally relevant dietary concentrations.

Rainbow trout were fed a diet containing 1.8, 18, or 90 pg/g 3H-2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) for up to 320 d. Concentrations of TCDD were determined in muscle, liver, and ovaries at 100, 150, 200, and 250 d. Concentrations of TCDD reached an apparent steady-state concentration in liver after 100 d of exposure, whereas concentrations in other tissues continued to increase until 150...

متن کامل

Rainbow Trout, Oncorhynchus mykiss

Rainbow trout are classified as Oncorhynchus mykiss, and as such belong to the same genus as Pacific salmon, and to the family Salmonidae, which includes Atlantic salmon (Salmo salar), various trout (Salvelinus sp.), Arctic char (Salvelinus alpinus), Arctic grayling (Thymallus arcticus) and whitefish (Coregonus sp.). Rainbow trout are native to areas around the North Pacific Ocean, from souther...

متن کامل

منابع من

با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

ذخیره در منابع من قبلا به منابع من ذحیره شده

{@ msg_add @}


عنوان ژورنال

دوره 17  شماره 3

صفحات  573- 584

تاریخ انتشار 2018-07-01

با دنبال کردن یک ژورنال هنگامی که شماره جدید این ژورنال منتشر می شود به شما از طریق ایمیل اطلاع داده می شود.

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023